JP 2011-526786 A 2011.10.20 10 (57)【要約】 本
Contents
1. 17 2011 526786 2011 10 20 H H H 0000 H E H 000000000002. in situl
3. 10 20 30 40 50 52 2011 526786 2011 10 20
4. 2011 526786 2011 10 20 46 l 10 20 l 000 l 30 40 50
5. H H 0000000000 50 Inn 10 20 30 40 50 2011 526786 2011 10 20 72 00 ID 10 20 ec DU l 30 40 50
6. 10 20 30 40 50 2011 526786 2011 10 20 60 10 20
7. 1000000000000000000 100000 00 00000 00 paid Ra 10 0000 00 00000000 oma 4000600 00 40 000 00 Da 2n 4 10 4 0000000 10 4 20 4 Inn 14 ius 10 10 4 1 4 1000000000000 id 6 IoD ia 140000 0000 0 E pl A 00 0000000 000 H 00 0 8 0 4000 B um ES 10 4 m 10 4 O 10 Mi a VV kumu walla Wu RIN 10000 1
8. 2011 526786 2011 10 20 47 10 20 JU DU l 30 40
9. CE C 1000 H 10 20 30 40 50 2011 526786 2011 10 20 84 10 20 30 40 50
10. 65 2011 526786 2011 10 20 H
11. 28 2011 526786 2011 10 20 000000 0000000000000 0000000 0000 0000000000 10 5 4 NH c 00 000 00000000 20 D 0000000 Doo 000 00000000 Dn n 00000000000 id ew i lt 5 N 7 00 Dn n 40 00 00000000 100000000000 29 2011 526786 2011 10 20
12. C 85 2011 526786 2011 10 20
13. H 00 00 IU IH l 00 DO 20 30 00010000 1000000600 1090 000000 iungit 40 50 36 2011 526786 2011 10 20 O LJ O L3 L3 I EP 10 1000000000 000000 000 20 000000000
14. 50 2011 526786 2011 10 20 15 10 20 l ID 30
15. 82 2011 526786 2011 10 20 10 20 30 40 50 83 2011 526786 2011 10 20
16. 40 00000 Ig 50 000 2011 526786 2011 10 20 13 l ID 10 20 30 40
17. AN NH Q 4 5 p hs d 10 00000000000000000 00000000 00 o Bop m po EE D 9B s p D ES 0000000000 c000 00000000000000000000000 20 N a N N i Zee 30 00000000000000 00000000000 NA 40 el l x d y T 50 2011 526786 2011 10 20 30 10 BS u 20 DU l
18. 10000000 000 10000000 10000000 10000000 ug 0000000 000000000000 00000000000 00000000000 ELA EL E Bp LOB TE PEDE 000 1000 1000000000 000 10 20 30 40 50 2011 526786 2011 10 20 4 10 20 l 000 n 30 40
19. 20 30 40 50 2011 526786 2011 10 20 57 10 20 30 l 40 l 50 n
20. 2011 526786 2011 10 20 2 r3 m C r1 ri C m
21. 10 1 0000D000 00000000000000 qu ma a 18 4 0000 10 4 EE E A 000 Mate 20 ina 24 000 0 30000 0000 2 0 400 O a V 5 D0 0000000 0 30 00 00000000000 0000 0 0000 0010 20 4 zd 10 4U 000000 kyl O 19 OSA Aaa aaa 00 008 30 0000000 10 PID 0 00 0000000000 00000 10 4 ka 10 4 al I 10 4 lina ll 1 40 gt 10 4 0 NE DER S 40 m S BUE TT Snell 0 0 B ag Bose Eb E Hl D D D 114 _ 2000 00 0000000000000000000000000
22. 50 2011 526786 2011 10 20 10 20 30 40 50 2011 526786
23. 1000 N 1 r NH N NH2 L F P D LD D n Im Im a 10 22 1 4 20 N o 4 RA e 30 OH NH N NA NH 40 Ea 0 000000000 LU 23 2011 526786 2011 10 20
24. 10 20 30 40 50 2011 526786 2011 10 20 75 10 20 30 l 40 50 2011 526786 2011 10 20 76
25. C o o 89 2011 526786 2011 10 20
26. 10 20 30 40 50 2011 526786 2011 10 20 onana 0O 10 20 30 40 50 2011 526786 2011 10 20 87 10
27. gt H 2011 526786 2011 10 20 45 10 20 30 40 n IU l n 50
28. 50 000 2011 526786 2011 10 20 5 10 20 30 40 50
29. 50 n 2011 526786 2011 10 20 14 000 10 l 20 30 40
30. H _ 5 H 10 20 30 40 50
31. 10 20 30 40 50 95 2011 526786 2011 10 20 unn E a x 6 s bg 10 20 30 40 50 96 2011 526786 2011 10 20
32. 10 20 30 40 50 2011 526786 2011 10 20 53 10 20 30 40 50 50 100 n 2011 526786 2011 10 20 54
33. 10 20 30 40 50 C C 77 2011 526786 2011 10 20
34. L3 J C 10 20 30 40 50 2011 526786 2011 10 20 97 10 20 30 40 50 l
35. 10 20 30 40 50 C C 71 2011 526786 2011 10 20
36. IB BE E Jp Th ED D A ER PT E E 1001000000 oa 000000 IUUUUUUUU j uU Hull HiuulIiu uiiliiiilil Y N u N T NH N OH 4 ai N N 1 0 10000 F uihil a E A E 000000000 0000000000 10 000000000 N Nm D 00 000 00 000 000000000 0 00000000 100 00000 00 10 0 1000000 00000000 Bs 10 20 30 40 40 2011 526786 2011 10 20 L1
37. 30 40 50 C 2011 526786 2011 10 20 101 10 n C 20 30 40 50
38. n 2011 526786 2011 10 20 10 20 30 40 50 2011 526786 2011 10 20 70
39. 10 20 30 40 50 8 2011 526786 2011 10 20 10 20 30 40 50 9 2011 526786 2011 10 20 H 10 20 30 40 50 2011 526786 2011 10 20 10 10 2
40. n n 56 2011 526786 2011 10 20 O 151
41. C 10 20 30 40 50 2011 526786 2011 10 20 68 10 20 30 40 50
42. 50 D 74 2011 526786 2011 10 20 y o E C ES a E gate H C C A
43. 000 C 40 50 2011 526786 2011 10 20 92 10 20 30 l
44. 10 20 30 40 50 63 2011 526786 2011 10 20 H H H H H H H ES 10 20 30 40 50 2011 526786 2011 10 20 64 10 20 30 40 50 C
45. 00000 NL V 3 F D LE D O n D D D T Data LR D D U Ph pa E 000000001000 D00 0 000 0000000 000000 0 00000000 0 O 00000000 00 000000 000000060 100000000000 IU LU O O O 0 000 00 00 4 0 0 40 UELUT sace DB B eem mag B e alri em s DOLCE 116 10 0 000 5 0
46. 40 50 l 2011 526786 2011 10 20 44 10 l 20 30 40 50
47. 00 27 2011 526786 2011 10 20 2 C F 2 O 10 Q u Do0 e Ten 0000000000 000000 00 20 SL N i Y W lt Q 7 30 000000 _ N N 40 N Y LA 50
48. 10 20 30 40 50 2011 526786 2011 10 20 66 10 20 100 30 40 50 C 67 2011 526786 2011 10 20 C
49. 40 50 scid l 2011 526786 2011 10 20 93 10 20 C 30 40 50
50. 40 50 2011 526786 2011 10 20 12 IU ID 10 20 30
51. 10 20 30 40 50 2011 526786 2011 10 20 103 10 IU ID ID DU 00000 l 20 104 2011 526786 2011 10 20 RI Sox1 19 1b 8 99 28 Zv H amp amp ateam PARE BENE Zv r5 T2T LZAT Wnt3ag A 67 42 8 12E 06 2 51 Zv k5 POFEVA WntSa 10619 100 00 8 2 08 100 00 20 05 08858 4848 45 66 1 60E 06 0 49 Zv k5 1 amp 59 0 19 4 62 06 1 43 ZvL k5 4 3348 31 52 2 05 0 07 2 5 1261 27 60 3 05E 06 0 94 2 5 181162 7171 67 53 2 04E
52. E E EET E E H 10 20 40 50 2011 526786 2011 10 20 81 10 20 30 40 50 C C
53. 10 20 30 40 50 2011 526786 2011 10 20 10 20 30 40 50 2011 526786 2011 10 20 91 10 20 30 l
54. 40 n 50 2011 526786 2011 10 20 16 10 20 30 40 n 50
55. 10 20 30 40 42 2011 526786 2011 10 20 SN N EE FL D jiundguauuuuldgasuululiuuili liiilLiiull 2011 526786 2011 10 20 43 n 10 20 30
56. 10 20 30 40 50 2011 526786 2011 10 20 38 1000 10 20 000 1000 h R l H IUD 30 40 D 39 2011 526786 2011 10 20
57. 8 0 80 DARE 4 70 50 17 ER si 60 60 17 8 8 50 50 40 5 40 2 30 20 ice 5 5 lt 10 t 220 se gt gt e 2 ES 2 lt Y xo gt N Qu Sy e 5 lt lt S S D S A SP Eg lt 59 Y S 5 i 8 0 UUU 7 0 5 17 8 0 eo 7 0 50 40 6 0 MSox17 5 3 5 3 0 de 5 0 S 3 20 i 2 40 10 5 E 9 oo Wa e lt 2 3 E T il E D s S gt amp n LE 10 I 0 0 gt gt o 2 y gt a m 2 SN UP S 59 Sd 9 N lt EP 80 14 B SOx17 E 7 0 7 8 12 E 3 M 10 60 ba os E 50 06 40 04 20 50 100 200 400 10 PeproTech GDF8 ng ml 0 0 Wnt3a lt E gt gt 9 e E ES ul SS lt 9 2 5 sE 49 SF 5 14 WSox1756Hi 12 4 000 3 dali w 104 1 4 4 485538 1 a 084 E 12 MSox17 5 amp 064 1 0 04 5 5 4 E 0 2 2 00 0 6 50 100 200 400 lt lt 04 Shenandoah ng ml 0 2 4 Wnt3a 0 0 Wnt3a AA Wnt3a
58. C 102 2011 526786 2011 10 20 00000000000 1 0000000600 MEE E nnnm pc
59. 15 141 RT PCR mi GAPDH AFP com cpx2 4 FOXA2 4 SATA6 Gsc KIT MIXL1 NANOG OTX2 POU5F1 SOX17 7 T AA 19 4 32 8 25 2 24 0 26 1 21 8 36 0 18 3 20 4 20 7 34 0 25 1 25 0 22 9 22 4 24 8 22 7 28 9 24 1 22 5 32 2 21 6 32 3 36 1 Wnt3a 18 2 40 0 23 5 220 24 1 20 9 40 0 17 1 22 0 18 7 33 4 22 9 23 1 21 8 22 8 23 8 22 9 27 7 227 199 31 0 21 2 81 2 34 1 AA 62181 20 1 40 0 245 23 3 26 0 20 7 35 9 18 2 22 0 20 1 35 1 25 6 24 7 22 9 20 8 25 5 19 7 27 8 248 223 32 8 22 2 330 83 8 AAT 180 18 4 342 23 5 216 25 9 21 4 352 17 2 220 18 9 34 0 24 0 23 1 22 6 22 5 24 8 226 27 9 237 203 32 1 21 4 32 0 32 7 20 1 35 4 24 4 24 9 26 3 20 8 40 0 17 9 17 7 20 5 32 8 25 6 25 1 22 7 21 3 26 0 22 6 29 5 24 1 22 9 32 1 22 3 33 0 29 6 12202 20 3 40 0 25 1 23 7 25 9 21 4 40 0 18 3 22 0 20 4 36 0 25 7 24 5 23 0 21 8 25 2 23 9 27 7 245 225 32 7 22 6 32 1 33 5 4 840 19 9 40 0 24 6 23 6 25 8 20 4 40 0 17 7 22 7 20 2 35 1 25 5 24 6 22 5 20 9 25 5 22 9 28 0 23 8 220 33 8 22 3 32 5 32 8 AAT gt 20 2 35 0 254 23 7 27 2 21 9 355 18 5 222 2
60. 58 2011 526786 2011 10 20 2 bi ucul l i ne C C C i nmn C H H H H H lt E CE H 2 10 20 30 40 50 59 2011 526786 2011 10 20 H
61. 2 HYPERLI NK http www mlli po C n re com catal ogue i ten 81 068 3 2011 526786 2011 10 20 73 10 100 20 30 40 000
62. 30 40 l 2011 526786 2011 10 20 e 10 c c F UDD Rm d is pup EN 20 N el Me NH 30 _ 00000 000 0 00 0000 _ EN xy 40 2 5 C J NH 50 2011 526786 2011 10 20 32
63. 10 20 C 30 40 50 l 100 2011 526786 2011 10 20 55 10 20 n 30 40 50
64. 6 000 5 000 4 000 3 000 2 000 1 000 5 0 156 0 313 0 625 1 25 25 5 10 AA 63 181 u M 6 000 5 000 4 000 3 000 2 000 1 000 0 313 0 625 1 25 25 5 10 19 38 0 0 078 0 156 4 6 000 5 000 4 000 3 000 2 000 1 000 5 0 156 0 313 0 625 1 25 25 5 10 AA 3815 202 u 3815 6 000 5 000 4 000 3 000 2 000 1 000 0 313 0 625 1 25 25 5 10 SER 28 34 38 0 0 078 0 156 m E 0 0 078 m ES 0 0 078 86 2 0 0 078 86 2 6 000 5 000 4 000 3 000 2 000 1 000 0 0 0 078 0 156 0 313 0 625 1 25 25 5 10 AA 40 6 000 5 000 4 000 3 000 2 000 1 000 0 156 0 313 0 625 1 25 25 5 10 AA GSK3i BIO 0 0 078 143 2011 526786 2011 10 20 2 fies 6 000 5 000 4 000 3 000 2 000 1 000 0 0 078 0 156 0 313 0 625 125 25 5 10 GDF8 2 181 u M 1818 6 000 5 000 4 000 3 000 2 000 1 000 5 0 0 078 0 156 0 313 0 625 1 25 25 5 10 GDF8 3815 1583 19 p M 1 Bf 2 6 000 5 0
65. 40 T N 0 g E gt de 50 34 2011 526786 A 2011 10 20 10 00000000 0000000 0000 100 0000000 1 00 000000 0000000 00000 CI N 10 N C D 20 0 0000000 0000 N 7 NH 30 v IS EN MN O 00 100000000000 0 40 0000 10000 0000 0000 000 0 10000000000 35 2011 526786 2011 10 20
66. 00000 X10 n5 1 78633 FIORI 9 latt E FIFCVAF 4273 33 70 4 75 07 17 49 FIFCVAE Wnt3a k 12676 100 00 2 72E 08 100 00 151434 AAT Wnt3a amp 8 FGF 50ng mL EGF 50ng mL 13317 105 06 2 01E 08 74 08 894 AAT Wnt3a S d FGF 50ng mL EGF 100ng mL 14189 111 93 2 1 08 73 90 088194 Wnt3a amp FGF 100ng mL EGF 50ng mL 12616 99 52 1 80E 08 66 21 18134 Wnt3a amp 5 FGF 100ng mL EGF 100ng mL 8269 65 23 1 13E 08 41 73 1688194 AATA Wnt3a EL ZU 11711 92 38 1 65 08 60 68 9818194 AAT Wnt3a amp 8 5L EGF 25ng mL 16052 126 63 2 14E 08 78 82 134 AAFS Wnt3ag t EL EGF 50ng mL 13593 107 23 1 94E 08 71 52 534 Wnt3a amp 8 EGF 100ng mL 13170 103 90 1 93E 08 71 04 6488194 AATA Wnt3ag FGF 25ng mL TEL 18433 145 41 2 49E 08 91 72 888194 AATA Wnt3a amp 4 FGF 25ng mL EGF 25ng mL 18841 148 63 2 60E 08 95 72 AAT E Wnt3ag t FGF 25ng 4 mL EGF 50ng 16232 128 05 2 30E 08 84 79 894 AAT Wnt3a AA FGF 25ng mL EGF 100ng mL 9309 73 44 1 08 51 00 984194 Wnt3a amp t FGF 50ng mL 12757 100 64 1 66 8 61 10 40483184 AAT Wnt3a t FGF 50ng mL EGF 25ng mL 17720 139 79 2 1 8 85 01 AAT Wnt3a FGF 50ng mL EGF 50ng mL 16331 128 83 2 26E 08 83 11 15434 AAT Wnt3ag 8 FGF 50ng mL
67. 30 40 C 50 100 AU t 61 2011 526786 2011 10 20 H H H H
68. 78 2011 526786 2011 10 20 LU Dd 00001041 IU Dd 10000000000 LU Dd Ul 10 20 30 40 50 2011 526786 2011 10 20 79 10 20 30 40 50 C MEET DU l 80 2011 526786 2011 10 20
69. 00060 10 20 30 40 2011 526786 2011 10 20 33 10 20 e C 30 00 D
70. 40 96 CFE ng ml 10 09 0 8 0 7 0 6 0 54 04 03 0 2 0 1 0 0 Ngn3 NKX6 1 EH PDX1 PIFIA GDF8AWnt3a E 582 40088 8 St 40 E82 C Ts GAPDH CDX MAFA NEUROG3 NKX6 1 PDX1 PTF1A GDF8 Wnt3a 321 19 0 248 310 24 1 24 3 22 5 25 4 GDF8 Wnt3a 2 18 6 24 3 29 8 23 1 23 9 22 0 247 GDF8 c 40 8321 19 3 24 6 31 3 23 9 24 0 22 3 25 0 GDF8 it AH 40 i312 18 7 24 9 30 6 22 9 23 7 21 8 24 5 CA Z FF ng ml GDF8A4E amp 40 1 88 m 12 5 AA Wnt3a 100 048 83 m2 3 GDF8AE amp 40 42 048 8 044 gt 149 2011 526786 2011 10 20 12 04 84 g g gt 64 41 MIEL CM AA Wnt3a GDF8 GDF8 Wnt3a DO 18 2 0 100 4 95 CXCRA 27 528 80 60 OD 490 40 4 20 GDF8 181 1 25uM 181 2 59 484 490 180 1 25 180 2 5 GDF8 4 6 1 5 490 GDF8 t3 19 2 5uM G F8 tt amp 12 34
71. HDI 00000000 H HL 1 D H s E m D s E cuero en 5 5 52 D 1000000000000 10 20 30 40 18 2011 526786 2011 10 20
72. 000 Q0000a p40 00 0 0 0 000 000000 000 0 ag 400 00 10 4 1 40000 6 0 50 19 2011 526786 2011 10 20 6 a ooi DICE e n 10 4 5 10 4 10 4 15 400 0000 10 11 13 E RES EL mE nmn 10 11 13 10 4 vai 10 4 m Hana lain 4 ZU p 4 00 00 10 4 n 000 00000 12 ip all 4 1 4 ag 4 0
73. 10 20 30 40 50 2011 526786 2011 10 20 99 10 20 30 40 50 n DU l ID 2011 526786 2011 10 20 100 10 20
74. 10 000000000 0000000 000 1000000000 20 100 0 00 0 000 000000 10000 0 000000000000 OI do B tt p 30 40 41 2011 526786 2011 10 20 7 N NH 0 mat L 00004 l XOU SA E 00 000 0 000 000 01 000 0 00 0 000000 e 5 0 00 000 H 3H ET a EE HH HEB EAE EL BEA
75. ERRORS RI lo Sox173838 7 ALE tamas BHAR 2 FGF PDGF A VEGF PDGF D A1E JL GDF8 114 2734 17 3 6 1 6 2 1 2 2169 13 7 3 77E 06 2 tate 3107 19 7 3 52E 06 1 2 2 17 3343 21 1 5 05 0 2 2 EGF FGF PDGF A VEGF PDGF D 1 E JL GDF8 8118 3034 19 2 2 37E 05 0 1 2 PDGF A VEGF PDGF D E L GDF8 1 8 119 2263 14 3 1 66E 06 0 6 2 PDGF A PDGF D GDF8 1771 11 2 5 572 06 2 0 2 EGF FGF PDGF A VEGF PDGF D 3 GDF8 24121 1136 252 1 79 7 6 3 2 EGF FGF PDGF A VEGF PDGF D AY TE L GDF8 124122 2021 12 8 2 07 7 3 1 FIFEVATE Wnt3a 6049 45 2 1 31 07 5 2 1 FOFEVA Wnt3a ZL 13392 100 0 2 50E 08 100 0 1 FGF PDGF A VEGF PDGF D GDF8 15878 118 6 2 67 106 5 1 FGF PDGF A VEGF PDGF D GDF8 2924 12714 94 9 2 46E 08 98 2 1 FGF PDGF A VEGF PDGF D AZ E L GDF8 124123 12165 90 8 2 15E 08 86 0 1 EGF FGF PDGF A VEGF PDGF D GDF8 821 12640 94 4 1 6 8 65 9 1 EGF VEGF PDGF D AY E L GDF8 163013 11491 85 8 1 61E 08 64 3 1 PDGF A VEGF PDGF D GDF8 11396 85 1 1 34E 08 53 4 1 FGF PDGF A VEGF PDG
76. 100 bg 94 2011 526786 2011 10 20
77. 2011 526786 A 2011 10 20 21 10 20 30 l 00 D 00 40 OH IUD Ll 1 50 22 2011 526786 2011 10 20
78. aniline pyri di notri azi 2011 526786 2011 10 20 H H 10 20 30 40 50 7 2011 526786 2011 10 20
79. 17 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 9 0 078 0 156 0 313 0 625 1 25 25 5 GDF8 5518 GSK3i BIO u M 181 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 0 078 0456 0 313 0 625 125 2 5 5 10 GDF8 12531 180 M 3818 5 17 o Sox1 738 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 0 0 078 0 156 0313 0 625 125 25 5 10 GDF8 388 1653 19 p M 3802 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 9 0 078 0 156 0 313 0 625 125 25 5 10 GDF8 3AM 1689 40 3888 5 17 Sox1 7585 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 0 0 078 0 156 0313 0 625 125 25 5 10 GDF8 5519 162 202 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 9 0 078 0 156 0 313 0 625 1 25 25 5 10 GDF8 151 34 381 141 2011 526786 2011 10 20 17 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 9 0 078 0 156 GDF8
80. 50 linage 48 2011 526786 2011 10 20 10 20 30 40 50 2011 526786 2011 10 20 49 l 10 20 30 40 50
81. 400 0 1458 m a E z 14 10 4 30 6 0 0 0 an 4 0 10 4 10 ME 3 m IN E 14 14 10 4 30 6 10 4 10 4 10 4 So 5 z 14 1448 nnnm n n n E 4 30 6 10 4 10 4 10 4 pu 10 4 OH Dani 4 N 10 20 30 40 50 20 2011 526786 2011 10 20 L3 L3 L3 oa N OH g E
82. ul 1 allcyl B 1 10 10 4 000 0 2000 2 0 4 104 10 4 0 000 000 DODO RETE su sm B B D 400 _ ics 40 000000000000 0 40000 0 0000 0000000000000 1000 HEB Make B 0 0 H Blog a DOPO muna D B nun pn g pami n D 11 400 000000000000000 10 20 30 40 50 25 2011 526786 2011 10 20
83. 18 5 229 23 3 Wnt3a 184 23 0 238 181 18 6 22 3 25 0 15 180 18 8 22 5 26 3 458 202 18 5 22 5 24 1 40 18 5 22 2 24 8 GSK3i IX bio 185 220 26 1 56 18 3 227 23 9 50 Ar 40 3 0 5 El 2 20 lt 10 0 0 y e S X lt n GAPDH INEUROG3 NKX6 1 19 0 237 233 27 4 237 296 Wnt3a 19 5 24 1 24 1 27 5 241 31 1 158 181 18 0 221 24 6 28 1 24 0 31 0 180 19 4 21 6 25 8 28 9 240 32 6 202 19 2 23 3 241 274 243 295 LE 19 2 22 9 25 3 28 7 29 1 35 312 GSK3i IXbio 18 0 227 27 7 32 0 265 337 tat 56 18 8 229 23 3 272 23 6 293 CDx2 PDX1 2 NEUROG3 23 NKX6 1 Bl 9 oo ma AA Wnt3al GAPDH 2 NEUROG3I 6 1 PIFIA 18 0 24 6 28 7 31 2 26 5 40 0 Wni3a 19 0 244 240 28 0 23 8 30 5 181 18 0 23 1 21 8 257 223 28 6 fest 180 19 5 239 23 7 277 238 307 Ace 19 19 1 22 9 26 4 30 2 25 1 326 202 19 0 24 5 22 2 25 9 23 0 27 9 129 40 19 0 236 227 25 6 234 284 GSK3i bio 19 1 23 0 23 8 279 242 29 5 56 18 9 24 0 23 0 26 3 23 5 27 7 5 0 40 8 30 5 lt
84. 2 e e x E 3 0 2 5 5 2 0 x 1 5 10 0 5 0 0 e 2 90909009 v EI 8 E E 4 x x x E e e SL e amp e amp E lt o 14 0 12 0 100 80 9 40 2 0 0 0 lt O o e e 100 80 60 40 20 0 gt AA Wnt3a IOGULUdU 2 5 2 0 m 1 5 5 E 10 m 0 5 0 0 s AA Wnt3a 60 55 504 454 H 404 2 354 2 30 254 2 0 4 lt 15 9 0 5 00 GDF8 t 2 202 97 100 2 30 60 y do 4 5 20 GDF8 Wnt3a 12811 2 0 15 ER 48 o E GDF8 t 35 40 GDF8 t 3 40 202 99 GDF8AVnt3a 8312 O 2 148 GCG EB ins 9 N S N 3 v 2 2 e Sa lt e gt lt lt lt g ES 202 2 NKX6 1 EN PDX1 PIFIA GDF8 t 4 202 248 88 m 235 GDF8 tt amp t 202 3 400882 2011 526786 2011 10 20 ng ml AA Wnt3a 95
85. __ gt ceri 7 catas 050 km mixL1 Fous 1 sox17 SOX7 Wnt3a 20 35 6 24 2 26 209 40 17 5 227 198 358 24 7 23 8 22 1 21 6 234 232 28 1 24 5 22 526 23 2 33 36 8 GDF8ZWwWnt3a 20 1 23 8 245 24 6 216 20 19 5 283 21 34 8 25 1 24 5 23 8 23 3 17 6 25 5 28 3 249 23 31 23 7 89 3 2 sino 1202 GDF8Z ito AETH NEURODI NEUROG3 NKX6 1 POU3F4 PTF1A 25 8 25 2 j 28 8 28 6 30 8 3 H 10 GDF8ZWnt3a GDF8 GSK3 BIO GDF8 AE GDF8 ite 202 GDF8 imao AIHA NEUROD1 NEUROG3 POU3F4 Vint3a 23 7 23 8 27 GDF8 Wnt3a 3 5 24 3 24 2 E 3 a 27 GDF8 GSK3 AER BIO GDF8 ite mie 20 4481202 3 3 3 GDF8 iama ALIAS NEUROD NEUROG3 4 AAZ Vint3a 21 7 27 28 4 GDF8 Wnt3a 22 6 28 3 28 9 GSK3 EE 2 BIO GDF8 42119 GDF8Z it 619202 GDF87 i amp t940 000000 30 128 2011 526786 2011 10 20
86. 00 50 DU DU l 0010 2011 526786 2011 000000000000 01104
87. 80000 AP GM LS SD SL 52 TZ UG ZM ZW EN BY KG KZ M RUT TM EP AT BE BG CH CY Z CK ES Fl FR FR HJ IE IS LT LT LU LV NQ PL SE 9 S K TR ON EF GA M G4 M SN TD TO AE AG AL AT BA EB BG BH BR BW BY BZ CA CH a QN CO G QJ CZ CE CK DM DO EG ES FI ME MG FR HJIDILINIS KE KG KM KN KP KR KZ LA LG LK LR LS LT LU LY MD ME M5 W W MZ NA NG NO NZ PE PG PH PL RQ RS RU SC SD SE SG SK SL SM ST SV SY TJ TM TN TR TT TZ UA UG 5 Z G W ZA MV 200000 73000 72000 1000 00 48065 AA90X BB31 BB34 BB40
88. 96 POIFEVAR E Wnt3a gA 7107 67 96 1 27 07 7 94 tal FPIFELALWnt3a 60E 08 Wnt3a EGF 27E 06 0817 AAT EGF FGF4 14E 06 AAT EGF FGF4 PDGF AB 05 817 Wnt3ag EGF FGF4 PDGF AB T A ZE JV 92E 06 0817 AAT Wnt3agh EGF PDGF A VEGF 14E 06 FGF4 PDGF A VEGF 29E 07 10 AAT EGF FGF4 PDGF A VEGF 97E 06 17 AAT Wnt3a t EGF FGF4 PDGF A Ay E L 1 45E 07 FL FIFEVAFRE Wnt3ag 1 27 07 uu TTULA ZWnt3a 1 60E 08 10488195 EGF 1 01E 07 2435 AAT Wnt3a EGF FGF4 1 36E 06 462 35 Wnt3a EGF FGF4 PDGF AB 1 28E 07 62935 Wnt3a EGF FGF4 PDGF AB A 7 JV 1 54E 06 68885 AAT Wnt3aS EGF PDGF A VEGF 1 23 07 115 935 AAT amp Wnt3a gA FGF4 PDGF A VEGF 3 92E 07 35 Wnt3a EGF FGF4 PDGF A VEGF 9 11E 07 amp 35 AAT EGF FGF4 PDGF A AZ 15457 147 80 6 89E 07 43 15 48929 Wnt3a 1971 18 84 1 44E 07 9 00 20 688329 AAT 8 EGF FGF4 7436 71 10 4 06 2 72 8029 AAT Wnt3a EGF FGF4 PDGF AB 89129 Wnt3a gA EGF FGF4 PDGF AB AZ E JV 083129 AAT Wnt3agh EGF PDGF A VEGF AAT amp Wnt3a gA FGF4 PDGF A VEGF 1529 AATA Wnt3a 3 EGF F
89. sus AAT tamo 18 5 258 34 9 22 2 aoo 223 aoo 29 7 34 1 28 1 29 1 30 3 33 6 32 9 40 0 248 30 3 31 3 35 6 35 3 35 4 AAT 4 18 5 24 9 30 1 34 6 22 0 40 0 21 5 40 0 30 2 349 27 8 340 31 0 31 8 32 6 40 0 34 9 40 0 261 32 4 400 352 34 9 AAT 13 8 27 0 30 3 33 7 22 7 357 225 28 4 34 4 27 8 30 7 28 6 29 8 32 8 35 0 1 5 304 239 29 7 31 2 85 0 31 9 40 0 GDFB 180 28 7 30 4 35 3 23 8 40 0 23 40 0 28 5 33 6 27 2 30 3 28 7 30 3 34 4 35 0 32 4 24 8 25 5 30 0 30 8 34 2 29 5 26 1 GDF8 Wnt3a 17 4 270 29 5 33 8 23 1 35 1 224 40 0 26 3 30 5 261 30 1 27 6 29 1 40 0 32 9 31 5 25 8 23 1 322 20 8 277 28 2 GDF8 AH180 18 8 27 3 30 6 32 4 22 8 34 9 227 40 0 29 5 35 9 27 7 300 27 5 29 0 35 4 32 6 31 2 40 0 237 316 30 8 34 6 34 4 100 GDF8 kamio 18 3 24 9 29 7 33 4 220 40 0 22 2 40 0 29 7 34 5 28 4 33 3 31 5 32 2 34 4 35 2 34 4 35 0 250 294 21 8 400 34 1 33 5 GDF8 amp 1 5202 18 7 278 30 4 32 8 23 8 40 0 22 9 34 6 28 5 34 2 27 6 29 6 27 1 28 4 35 7 30 5 4 31 8 230 30 2 316 30 9 34 0 GDF8 Amao 18 4 27 7 30 1 32 5 23 0 35 1 224 40 0 29 1 34 3 27 5 27 8 28 8 35 3 32 9 31 5 40 0 28 2 292 30 5
90. 2 100 000 000 50 000 000 0 0 0 078 0 156 0313 0 625 125 25 5 10 AA 1583 40 u 18 8 300 000 000 250 000 000 200 000 000 8 B C 150 000 000 5 5 5 100 000 000 50 000 000 0 0 0 078 0 156 0313 0 625 125 25 5 10 380 12 18 300 000 000 250 000 000 200 000 000 150 000 000 5 8 hs 100 000 000 5 50 000 000 o 0 078 0 156 0 313 0 625 1 25 AA 3815 162 180 38M 2 5 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 j 0 078 0 156 0 313 0 625 1 25 25 AA 15 202 u M 388 139 2011 526786 2011 10 20 17 Sox1 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 9 0 078 0 156 0 313 0 625 AA 158 40 p M 1 25 25 5 10 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 9 0 078 0 156 0 313 0 625 25 5 10 AA GSK3i BIO p M 3813 1 25 Sox1 755 e Sox1758FF 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 9 0 078 0 156 0 313 0 625 GAR 83 34 SAM 1 25 25 5 10 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000
91. 7 50 95 Zv k2 15855106 4211 37 06 3 94ET O7 40 Zv k2 15893107 2853 25 11 1 78 07 18 04 106 00 00 ERRORE J P 2011 526786 A 2011 10 20 RI RO 17 Zv H amp S altxa 90 Zv k2 3779 33 27 2 07 24 26 270 2 2869 25 26 2 07 20 71 260 2 1556593110 4398 38 71 2 53E 07 25 65 20 63 91 17 1 17 07 5 89 71 03 244 13 1 98E 08 100 00 2 63 1859111 240 04 5 7 26 90 70 63 138112 5357 188 66 3 52E 07 17 74 2 3 211 37 8 55E 07 43 11 BL k3 116 49 3 85E 07 19 44 71 03 Zv ks Zv ta 22 63 215 81 5 53 07 27 91 Zv F3 124119 4184 147 35 3 41E 07 17 21 71 63 87 66 2 87E 07 14 49 20 03 15833121 4985 175 54 3 94E 07 19 87 7 3 146 17 4 07 20 32 70 63 14122 6407 225 61 4 15E 07 20 95 2 3 157 24 5 35E 07 26 99 Zv ka 155 58 4 67E 4 07 23 55 20 03 25 6367 224 23 5 73 07 28 93 7 3 216 82 7 47 07 37 70 2712 63 196 97 5 61 07 28 28 Zv k3 146 50 4 91E 07 24 77 2 03 13129 3778 133 3 54E 07 17 88 7 63 1E amp 93130 4357 15E 07 JZv F3 1859131 6135 4 28E 07 27 63 1441132 4421 4 58E 07 2 797 110 9 79E 07 100 210 4 169134 5277 139 62 3 7 35 04 Zv k4 149 67 3 38E
92. 0 000 00 00 102 0 ne oe EPOR BU Ti 1660 D 6 IDD 100000 1000000000 50 24 2011 526786 2011 10 20
93. 2011 526786 2011 10 20 19 PT 3352011 526786 P2011 526786A UJARE sEFR234E10F20E 2011 10 20 51 Int Cl 2 12 5 0735 2010 01 C12N 5 00 202 48065 ARR 7 RAR 155 HiRes 4 882011 516798 P2011 516798 71 HERA 509087759 6 22 3E BE214E6H 2009 6 30 AY A 5 EI 3E pR237E2H 24A 2011 2 24 6 57 PCT US2009 049183 TAVARES ARA MM WO2010 002846 ARTIPA 51780 EE EZ 2010 1 7 0 850 14 110000741 AH 80 2008 6 30 RRT EA 3 US 61 076 900 2 3E 30H 2008 6 30 72 UD 27072 y30Z 5 BH 2 2Y r748085 21 86 85 86 87 87 31 61 076 908 32 33 31 32 5 1 33 31 9 US 99 BULHEXGRERE 61 076 915 32 3E BE 204E6H 30H 2008 6 30 33 05 lt 54 SERENO 520000 Figure 000000000000000000000000 0000000000000000000000000 NETT iG 0000000000 00000000000000 0000000000000000000000000 8 10000000000000000000
94. EGF FGF4 WntSa 14158 185 07 2 40 215 35 2 10ng mLDAA FGF4 Wnt3a 13323 174 15 2 19E 08 196 86 2 10ng mMLMAA 8194 EGF Wnt3a 14527 189 89 2 28E 08 204 84 2 1Ong mLOAA 34 Wnt3a 3589 46 91 7 2 7 63 08 EGF FGF4 PDGF A VEGF PDGF D 2 10ng mLDAA 49499 AYE J LA4GDF8 Wnt3a 5738 75 00 2 14 07 19 24 2 lOng mLOAA 9 2 82E 06 2 53 2 10ng mLOAA 12939 EGF FGF4 PDGF A Wnt3a 2879 3 61 06 3 24 2 lOng mLOAA EGF FGF4 PDGF AB Wnt3a 2989 39 07 1 78 04 0 02 2 10ng mLQAA 083199 EGF FGF4 Wnt3a 3 93E 06 3 53 2 1O0ng mLOAA 8139 FGF44 Wnt3a 4 46 06 4 01 2 lOng mLOAA 42 39 EGF Wnt3a 4 54E 06 4 08 2 1Ong mLOAA 48139 Wnt3a 2 37E 06 2 13 EGF FGF4 PDGF A VEGF PDGF D 2 10ng mMLQAA 185337 LYE IL GDF8 Wnt3a 1 21E 07 10 82 2 10ng mLQAA 24137 EGF FGF4 PDGF AB VEGF Wnt3a 5 16 05 0 46 2 10ng mLQAA 6658137 EGF FGF4 PDGF A 4 E JL Wnt3a 9 23E 05 0 83 2 1O0ng mLOAA 6850137 EGF FGF4 PDGF AB Wnt3a 6 15 05 0 55 2 10ng mLOAA EGF FGF4 Wnt3a 5 60E 05 0 50 2 lOng mLOAA 89037 1 885406 1 69 2 10ng mLDAA 37 EGF Wnt3a 1431 18 70 2 75E 06 2 47 2 10ng mLQAA 37 Wnt3a 791 10 34 2 99E 06 2 68 10 20 123 2011 526786 2011 10 20
95. 2 1E 05 0 1 1 123142 60 1 2 4E 05 0 1 1 857 8 4 7 1E 05 0 3 1 GDF8T 8110 48 4 6 05 0 3 GDF8 69 0 5 9 04 0 0 1 68937 0 0 1 4 06 0 6 GDF8 40480145 61 2 4 5E 05 0 2 GDF8 408336 GSKE RIE 102 4 1 8E 05 0 1 GDF8 108339 49 4 4 2E 04 0 0 temz 90 7 2 05 1 GDFS T 13235 91 4 3 8E 05 0 2 GDF8 13926 96 2 2 6E 08 111 8 1 GDF8 8145 65 9 1 1E 08 47 9 1 GDF8 GSKiRTE 36 6 7 0 07 30 4 GDF8 32 0 6 6E 07 28 6 1 GDF8 108316 35 4 5 8E 07 25 2 GDF8 0831498 Tri 46 2 5 4E 07 23 4 GDF8 15142 4 39 3 4 9E 07 21 2 GDF8 68338 GSKIEARTE 34 7 4 7 07 20 4 GDF8 158529 33 3 4 07 19 4 GDF8 L34941 GSKj3ERTE 30 8 3 4E 07 14 8 1 GDF8 n a n a 19 7 2 2E 07 9 4 1 GDF8 6257 14 6 1 1E 07 4 8 1 GDF8 158511 210 1 4 4 9E 05 0 2 1 GDF8 1080144 226 1 6 9 5E 05 0 4 1 GDF8 62912 31 0 2 1 SEX 06 0 6 131 2011 526786 2011 10 20 18 7 Zv k 3E sp cv 9 sp cv fe TUET LDAT 1
96. GSK3i BIO u M SEM 0 313 0 625 125 25 5 10 6 000 5 000 4 000 3 000 2 000 1 000 DI 0 0 078 0 156 0 313 0 625 1 25 25 5 10 815 181 u M 1519 6 000 5 000 4 000 3 000 E 2 000 1 000 6 000 5 000 4 000 3 000 2 000 1 000 0 0 078 0 156 0 313 0 625 1 25 25 5 10 AA 88 11659 180 u M 1808 6 000 5 000 4 000 3 000 4 2 900 1 000 0 0 078 0 156 0 313 0 625 1 25 25 5 10 388 123 19 u 1879 6 000 5 000 4 000 3 000 2 000 1 000 0 0 078 0 156 0 313 0 625 1 25 2 5 5 10 380 4 1539 40 EM 6 000 5 000 4 000 3 000 4 2 000 1 000 0 0 078 0 156 0 313 0 625 1 25 25 5 10 38 202 1815 6 000 5 000 4 000 3 000 2 000 1 000 0 0 078 0 156 0 313 0 625 1 25 25 5 10 AA 3813 145 34 M 1858 142 2011 526786 2011 10 20 3 6 000 5 000 4 000 3 000 2 000 1 000 o 0 156 0 313 0 625 1 25 25 5 10 AA GSK3i u M 1818 6 000 5 000 4 000 3 000 2 000 1 000 0 313 0 625 1 25 25 5 10 15 180 3815 0 0 078 0 156
97. Sox1756H5 ite ES sk Blt Rm 8896 Wnt3a 13392 100 0 2 50E 08 100 0 EGF FGF PDGF A PDGF D A2E JL GDF8 820 9434 70 4 1 48E 08 59 1 EGF FGF PDGF A VEGF PDGF D A 7E JL GDF8 2917 7988 59 6 1 13E 08 45 0 1 PME 8303 62 0 9 20E 07 36 7 1 EGF FGF PDGF A VEGF PDGF D GDF8 158 13 7045 52 6 7 2 07 28 8 1 PDGF A VEGF PDGF D E L GDF8 7799 58 2 6 82E 07 27 2 1 EGF FGF PDGF A VEGF PDGF D AY TE L GDF8 1094192 5886 44 0 5 63E 07 22 5 1 EGF FGF PDGF A VEGF PDGF D A E L GDF8 ite 093 5463 40 8 4 38E 07 17 5 1 FGF PDGF A VEGF PDGF D Av E L GDFS 94 5100 38 1 4 18 07 16 7 1 FGF PDGF A VEGF PDGF D AL E L GDF8 083195 4510 33 7 3 32 07 13 3 1 FGF PDGF A VEGF PDGF D E L GDF8 993196 4570 34 1 3 09E 07 12 3 1 FGF PDGF A VEGF PDGF D GDF8 983197 4561 34 1 2 15 07 8 6 1 FGF PDGF A VEGF PDGF D A E JL GDF8 4494198 3176 23 7 9 86E 06 3 9 1 EGF FGF PDGF A VEGF PDGF D GDF8 984199 1209 9 0 1 56E 07 6 2 2 FIFUCVAT Wnt3a 15494 98 0 1 25E 07 4 4 2 FIFEYVA Wnt3a ZU 15807 100 0 2 86E 08 100 0 2 FGF PDGF A VEGF PDGF D 08118 8742 55 3 1 01E 08 35 4 2 FGF PDGF A VEGF PDGF D 55
98. Wnt3a tA 15494 98 0 1 25E 07 4 4 ZL TZTULA Wnt3a 15807 100 0 2 86 8 100 0 1818 PDGF A VEGF PDGF D E JL GDF8 8742 55 3 1 01E 08 35 4 000000 10 20 30 40 000 00 113 2011 526786 2011 10 20 ERRORS RA E Sox175H lE ERRARE BERTA 96 14 PDGF A VEGF PDGF D A EJ GDF8 8464 53 5 8 07 29 1 62915 FGF PDGF A VEGF PDGF D 2 GDF8 7234 45 8 7 95E 07 27 8 12100 EGF PDGF A VEGF PDGF D AvE L GDF8 6805 43 0 5 88E 07 20 6 EGF FGF PDGF A VEGF PDGF D 2 GDF8 5668 35 9 5 34 07 18 7 1585102 EGF FGF PDGF A VEGF PDGF D E L GDF8 6195 39 2 5 29 07 18 5 158 7103 FGF PDGF A VEGF PDGF D GDF8 7545 47 7 5 13 07 18 0 154104 FGF PDGF A PDGF D GDF8 4757 30 1 4 58E 07 16 0 15891105 EGF FGF PDGF A VEGF PDGF D E L GDF8 6285 39 8 4 29E 07 15 0 62106 EGF FGF VEGF PDGF D 2 GDF8 5622 35 6 2 86E 07 10 0 181107 PDGF A PDGF D Av E L GDF8 3951 25 0 1 72 07 6 0 40981108 PDGF A VEGF PDGF D 2 GDF8 3226 20 4 1 58E 07 5 5 1629109 PDGF A VEG
99. lt GAPDH GCG INS NIOQ 2 NKX8 1 PDX1 17 4 18 1 141 25 9 26 6 26 6 25 5 Wnt3a 17 8 17 7 17 5 25 6 27 0 26 5 25 6 181 18 0 145 148 234 25 0 24 4 23 0 180 180 14 2 144 23 2 25 1 23 8 229 19 18 3 16 2 15 6 24 3 286 252 24 2 202 18 7 15 7 15 8 23 8 24 8 25 2 23 2 158 40 18 1 14 8 14 8 23 2 25 0 24 1 229 GSK3i bio 18 4 15 2 15 5 242 26 7 24 8 240 458 56 18 0 16 0 16 0 24 4 25 9 254 240 2 5 2 0 5 x 15 m 10 0 5 0 0 147 2 NEUROG3 NKX6 1 ES PDX GCG INS NKX2 2 NKX6 1 6 PDX1 2011 526786 2011 10 20 O eco 50 O INS m 2 E 40 NKX6 1 6 R 30 PDX1 2 2 0 N lt lt GAPDH GCG INS 2 NKX6 1 PDXt tal 18 3 14 1 149 23 7 25 5 24 0 23 2 Wnt3a 18 0 14 0 14 6 234 28 1 23 7 23 3 181 18 0 14 3 14 5 23 3 26 1 24 0 23 1 180 18 0 14 9 14 7 23 8 26 5 24 3 23 2 19 17 9 27 0 26 9 28 0 258 28 6 294 202 18 6 14 9 15 0 23 7 25 8 24 7 23 6 40 18 3 146 14 7 23 5 26 1 24 4 232 GSK3i IX bio 18 3 15 1 15 1 242 27 8 24 9 23 8 56 18 2 14 4 14 8 23 6 25 9 24 1 23 3 U 47 5 E ins 15 0 125 x 10 0 dn 7 5 8 50 2 5 0 0 y gt D gt S gt 9 S EJ
100. 242 290 30 1 31 0 20 s 31 9 GDF8 amp 202 18 7 19 6 28 3 23 1 24 0 15 7 21 9 15 8 23 5 32 8 23 3 22 3 27 8 23 8 31 3 24 8 23 1 25 2 29 0 29 5 27 1 20 8 30 9 GDF8 12240 18 1 18 7 27 9 22 3 23 0 14 8 21 1 14 8 22 6 31 5 22 5 21 5 2238 23 2 33 2 25 0 26 6 24 1 22d 273 28 9 27 4 20 4 32 0 ADEESBIO 18 4 17 1 27 6 23 2 228 15 2 205 15 5 23 1 32 6 22 6 22 1 2 24 2 35 0 26 7 27 3 24 8 240 281 30 1 28 5 19 3 32 4 GDF8 129206 18 0 20 0 27 9 23 4 24 0 16 0 267 16 0 23 6 32 1 22 9 22 8 28 5 24 4 30 5 25 9 27 5 25 4 24 0 29 7 28 3 28 0 20 8 23 9 16 LEES EERI 1606 GSKGRHIR TE 018318 5 689157 L64941 42 L34910 088194 811 168 44 412 2445 RE 130 2011 526786 2011 10 20 17 50 17 L A 31085 3E HEAR RO E 1 FIFCVATE n a 9809 67 8 4 0 05 0 2 1 FIFUVA Wnt3a n a 14476 100 0 2 3E 08 100 0 1 GDF8 T 565 3 9 1 1E 06 0 5 1 GDF8 2 15444 GSKJEHRTE 14 0 1 1 1 06 0 5 1 GDF8 15143 59 5
101. 28 9 237 24 5 21 8 22 3 21 7 24 3 30 3 23 5 22 9 22 2 23 4 33 4 25 9 24 8 26 5 230 29 5 26 1 SI 22 4 34 1 GDF8 121940 19 0 21 4 29 0 234 23 6 21 0 22 0 21 5 23 8 30 2 23 3 23 2 227 23 7 33 2 26 6 24 8 26 1 234 29 3 26 4 28 4 22 7 32 6 GDF8 GSK3 4 19 1 19 4 29 1 24 3 23 0 21 5 21 4 21 8 246 31 0 24 0 23 8 24 2 33 3 27 9 26 1 27 0 242 300 27 8 29 5 23 4 32 7 GDF8 1206 18 9 21 8 26 9 24 4 24 0 22 6 22 3 22 7 24 9 30 9 24 2 23 7 230 24 0 35 3 26 3 25 5 27 2 23 5 31 4 26 9 27 7 24 0 28 9 T 5 RT PCR GAPDH ALB AMY2A ARX CDx2 ece INS ISL1 NEUROD1 NEUROG3 2 2 5 1 PDX1 POUSF4 PTF1A 557 zci AA 18 3 20 2 276 220 23 4 14 1 21 3 14 9 224 31 8 22 2 22 8 281 23 7 34 1 25 5 27 8 24 0 232 301 28 8 27 3 19 6 34 4 AAT Wnt3a 18 0 20 0 27 7 21 9 23 1 14 0 20 9 14 6 22 3 31 6 22 0 21 6 280 28 4 34 8 26 1 27 9 28 7 23 3 296 28 8 27 5 19 4 32 5 AAT 1629181 180 188 276 22 0 22 143 209 2 1 31 4 2 2 215 28 23 3 322 261 26 8 240 231 27 6 8 8 280 18 8 312 AAT t amp mo 18 0 18 8 27 6 22 4 22 9 14 9 21 0 14 7 22 4 31 9 22 6 21 7 29 5 23 8 30 2 26 5 27 2 24 3 23 2 26 7 29 0 28 7 18 7 30 3 AAT Stato 17 9 23 8
102. 00 000000 1000 0000 Ver E 10 4 nd B 1 4 10 4 sE 10 4 E 10 000000 000 10 10 4 10 4 10 4 10 4 ve 14 n 14 10 4 10 4 4 30 6 4 l 1 1 14 1 30 10 400000 CANS D 000 0000 00 NA S m VA P xd 100000000 10 20 30 50 2011 526786 2011 10 20 26 ID DU l n 10 l 20 l nnnn nnnn 30 40 N
103. 17 31 2 208 209 21 2 18 4 35 3 15 5 19 4 17 2 29 7 21 1 20 8 19 6 20 21 8 21 5 25 21 1 19 3 27 9 21 8 31 1 30 3 AIRS pam GAPDH ALB AMY2A ARX CDX2 GCG HNF4 INS ISL1 MAFA MAFB NEUROD1 NEUROG3 NKX2 2 NKX2 5 NKX6 1 PAX4 PDX1 4 PTF1A SST ZIG1 ba 18 7 23 1 30 2 8 22 4 34 1 21 1 34 9 27 9 35 1 26 8 30 6 28 29 3 33 1 38 8 30 6 36 256 28 3 29 5 38 4 30 3 32 7 18 4 23 1 29 7 9 22 5 34 5 21 1 34 7 27 4 34 6 269 30 5 28 36 4 36 31 1 335 25 4 30 4 30 1 38 5 27 9 32 4 ossa 18 4 23 3 29 7 32 7 22 5 36 6 21 1 38 1 27 5 342 26 8 333 31 32 2 33 8 37 8 33 2 365 26 3 28 1 31 1 36 4 27 6 33 2 Lexi 18 2 23 5 29 7 1 6 22 6 36 3 21 2 35 7 27 3 34 4 27 30 6 27 9 33 8 38 2 30 8 32 7 25 28 9 30 4 348 27 5 32 7 2E GAPDH ALB AMY2A ARX HNF4 INS ISL1 MAFA MAFB NEUROD1 NEUROG3 NKX2 2 NKX2 5 NKX6 1 PDX1 PECAMI POUSFA PTF1A SST A 18 3 18 3 27 1 23 8 21 7 19 5 20 6 20 5 23 3 31 23 8 23 27 8 24 6 31 5 B E 27 5 26 4 24 1 26 9 27 6 40 31 4 nd 18 9 19 3 27 7 24 1 g 19 9 21 2 20 7 23 7 BUE 24 2 23 5 27 8 25 35 4 31 5 27 7 26 3 24 5 29 5 29 3 38 2 24 9 31 4 18 9 18 9 27 1 25 22 3 21 5 21 22 2 24 4 31 3 246 246 29 7 25 8 31 6 29 6 27 9 264 23 9 27 5 28
104. 17E 06 3 65 1 10ng mLQAA itens FGF4 Wnt3a 7 54 06 6 59 1 lO0ng mLOAA 1525935 EGF Wnt3a 7 82 06 6 84 1 10ng mLQAA 35 Wnt3a 7 61 06 6 66 EGF FGF4 PDGF A VEGF PDGF D 1 10ng mLDAA 1 29 LU E IL4GDF8 Wnt3a 2 41 07 21 10 1 lO0ng mLOAA 483129 EGF FGF4 PDGF AB VEGF Wnt3a 5 59E 06 4 88 1 1O0ng mLOAA 98129 EGF FGF4 F PDGF A 2 41 07 21 11 1 10ng mMLMAA 84129 EGF FGF4 PDGF AB Wnt3a 2 27E 07 19 89 1 10ng MLMAA 2129 EGF FGF4 Wnt3a 1 98E 06 1 73 1 10ng mLDAA 1038129 FGF4 Wnt3a 6 98E 06 6 10 1 10ngmLMAA 158129 EGF Wnt3a 4 86E 06 4 25 1 10ng mMLMAA 12929 Wnt3a 4 89E 06 4 28 2 IL 3 35 06 3 01 122 2011 526786 2011 10 20 RO Sox173618 JL TUT VA RAF FRE pesos 2 10ng mMLQAA 5177 67 67 3 89E 06 3 49 2 100ng mLOAA 7650 100 00 1 11E 08 100 00 EGF FGF4 PDGF A VEGF PDGF D 2 1O0ng mLOAA ite 2 1 GDF84 Wnt3a 18362 240 02 3 45E 08 309 74 2 10ng mMLQAA EGF FGF4 PDGF AB VEGF Wnt3a 15574 203 58 2 59E 08 232 70 2 10ng mLDAA 12434 EGF FGF44 PDGF A 17890 233 85 2 88E 08 258 30 2 10ng mMLQAA EGF FGF4 PDGF AB Wnt3a 17875 233 65 2 68E 08 241 07 2 l0ngZmL AA
105. 22 2 21 8 23 5 25 3 31 5 24 7 24 7 25 3 25 0 32 2 28 7 27 1 28 2 25 1 30 3 285 31 2 25 6 32 7 GSK3 3 D 19 1 2 TR 22 7 25 2 211 26 3 26 5 33 0 254 26 3 21 1 PER 315 32 8 28 4 30 0 26 5 29 6 30 5 33 7 272 32 4 tE 51206 18 8 20 9 284 23 3 229 212 21 7 22 8 24 0 30 4 23 5 233 23 3 23 8 31 9 27 2 25 5 26 3 23 6 297 266 293 236 az 2 18 0 25 5 29 1 29 8 24 6 31 3 238 30 9 27 6 32 8 24 1 292 28 7 29 0 32 8 31 2 1 1 27 9 265 294 29 4 40 0 260 22 8 129 2011 526786 2011 10 20 ERRORS GDF8 Wnt3a 19 0 24 3 29 0 25 4 244 27 7 23 0 25 5 25 4 32 7 25 1 25 4 24 0 25 0 33 3 28 0 26 7 28 8 238 334 27 7 30 5 24 1 29 0 GDF8 84181 18 0 22 8 28 1 23 5 23 1 24 6 21 4 22 7 23 8 30 6 23 0 22 9 21 8 23 2 32 1 25 7 24 2 27 0 223 276 25 4 28 6 24 2 31 6 GDF8 16840180 19 5 24 0 223 244 23 9 25 7 22 5 24 5 247 31 4 24 4 24 3 23 7 24 4 83 5 27 7 26 0 28 3 238 26 9 30 7 26 3 83 4 GDF8 1683319 19 1 226 287 25 5 229 26 7 22 1 27 2 35 8 32 6 25 5 26 2 26 4 25 6 34 0 30 2 28 2 251 302 29 2 32 6 27 9 81 1 GDF84 8202 19 0 22
106. Wnt3a 8 EGF FGF4 20221 107 97 2 83E 07 15 27 1689339 AAT Wnt3a amp EGF FGF4 17095 91 28 2 82 07 15 19 1589537 AAT Wnt3a d EGF FGF4 15605 83 32 2 67E 07 14 37 LS 35 8 f EGF FGF4 23823 127 20 2 54E 07 13 69 9580 AAT Wnt3a amp EGF FGF4 19864 106 07 2 07 12 54 28141 AAT Wnt3Sa EGF FGF4 17719 94 61 2 24E 07 12 04 1t amp 530 Wnt3a EGF FGF4 18063 96 45 2 18E 07 11 73 08150 AA Wnt3a EGF FGF4 16833 89 88 2 16 07 11 63 2 144 AAT EGF FGF4 17100 91 31 2 04E 07 11 01 158453104 AAT Wnt3a amp EGF FGF4 17863 95 38 1 89E 07 10 19 15845142 AAT Wnt3a amp EGF FGF4 18955 101 21 1 84 07 9 90 154110 AAT EGF FGF4 17534 93 62 1 76 07 9 45 12478 EGF FGF4 17703 94 52 1 71E 07 9 23 154133 AAT Wnt3ag f EGF FGF4 16521 88 22 1 67 07 8 97 688187 AAT amp Wnt3a EGF FGF4 16495 88 07 1 55E 07 8 33 95 EGF FGF4 16900 90 24 1 43E 07 7 72 136 AAT Wnt3as t EGF FGF4 19167 102 34 7 91E 06 4 26 105 EGF FGF4 15217 81 25 7 45E 06 4 01 15835134 Wnt3a amp EGF FGF4 17208 91 88 7 40 06 3 99 L 7138 AAT WntSa amp EGF FGF4 16695 89 14 6 65 06 3 58 1585589 Wnt3a amp EGF FGF4 14652 78 24 3 89E 06 2 10 143290 WntSa amp EGF FGF
107. Wnt3a amp 103 2 3 752 07 00E 00 00 10 9 1 FIFEVA Wnt3a AL 100 0 3 45E 08 7 16E 07 20 75 100 0 1 GDF8 1299206 137 1 5 19E 08 4 41 07 8 51 150 3 1 GDF8 1580207 128 1 2 54E 08 69E 07 22 41 73 6 1 GDF8 83 8 2 40 08 6 34E 07 26 44 69 5 1 GDF8 13423 58 4 1 16 08 9 07 06 7 80 33 7 1 GDF8 24170 43 0 9 44 07 6 98E 07 73 93 27 4 1 GDF8 14208 32 3 2 26E 07 2 15E 07 94 96 23 6 1 GDF8 209 32 2 3 97 07 1 61E 07 40 56 TiS 1 GDF8 16632 28 9 3 63E 07 3 27 07 90 14 10 5 1 GDF8 43190 3152 3 47 7 1 04E 07 30 03 10 1 1 GDF8 1580223 24 7 3 45 07 2 07 79 43 10 0 1 GDF8 342 30 2 2 95E 07 2 81E 06 9 50 8 6 1 GDF8 1 210 17 6 2 90E 07 2 45E 07 84 73 8 4 1 GDF8 1081124 38 4 2 64E 07 1 66 07 62 95 7 7 1 GDF8 124211 34 4 2 61E 07 5 02E 06 19 23 7 6 1 GDF8 1580212 36 5 2 54 07 1 18 07 46 53 7 4 GDF8 84224 5188 761 14 67 34 6 2 46E 07 8 26 06 33 56 7 1 1 GDF8 12 225 29 5 2 45 07 2 65E 07 108 19 7 1 1 GDF8 13 20 8 2 44E 07 2 30E 07 94 13 7 1 1 GDF8 24213 8 4 2 07 1 97 07 95 03 6 0 1 GDF8 52 82 9 1 99E 07 6 90E 06 34 67 5 8 1 GDF8 8214 22 3 1 93E 07 1 39E 07 72 18 5 6 1 GDF8 5451 28 6 1 70 07 1 10E 07 64 86 4 9 1 GDF8 1091126 82 6 1 65 07 5 93E 06 36 02 4 8 1 GDF8 12226 24 1 1 59E 07 4 96E 06 31 21 4 6 1 GDF8 124215 28 8 1 45 07 2 69E 06 18 53 4 2 1 GDF8 1531 24 1 1 36E 07 4 63E 06 34
108. 04 0 2 0 AA Wnt3a GDF8 Wnt3a GDF8 34 GDF8 6841 56 16 a 1 385 14 Cdx2 Cdx2 5Pdx 10 Ec 9 10 6 08 8 2 06 04 02 0 0 GDF8 Wnt3a 283 34 GDF8 1689 56 2 0 rs Pdx1 E 1 6 14 P 19 10 508 06 5 02 0 0 AA Wnt3a GDF8 Wntda GDF8 168 34 GDF8 158 56 JP 2011 526786 A 2011 10 20 7FIFCVA GDF8 170 3 0 5 0 0 0 5 1 0 1 5 2 0 2 5 3 0 3 5 4 0 Log ng ml 80 74 0 70 6 70 57 3 56 4 ENE 40 30 20 10 AAWnt3a GDF8 Wntsa GDF8 34 GDF8 56 2 5 m nde INS GCG R BGCGI A 3 20 0 5 0 0 AA Wnt3a GDF8 Wnt3a GDF8 8 1 34 GDF8 5 56 80 m sScx173 3R 3 0 2 0 1 0 0 0 x 9 z N lt lt 8 0 70 6 0 5 0 40 3 0 2 0 1 0 2 LE D 982 B Sox 175681 rx AAZWnt3aXtBE1 136 2011 526786 2011 10 20
109. 06 0 63 70 05 Jv ke FIFEVAFE WNt828A ZvL k6 FPOIFELVA Wnt3a 100 00 3 14E 08 100 00 2 6 4090163 1917 19 36 4 75E 05 15 Zv ke it amp tn26 6 Zv F6 1828 4121 41 62 1 89E 06 Zv k6 1585529 5164 52 15 1 66E 06 JZv k6 12430 4726 47 73 1 23E 06 L F7 FIFCVAF Wnt3a 4 87E 06 L PF7 FIFCLA Wnt3a 4 92E 08 270 7 6231 3 45E 06 L E7 1232 14655 73 04 3 03E 06 0 62 Zv k7 62 33 13891 69 23 8 11E 06 1 65 Zv k7 108 8194 58 18 2 24E 06 0 46 20 07 1585535 15379 76 65 7 30E 06 1 48 20 07 15 9136 8356 41 65 4 57E 06 93 2 Wna fA 6868 36 97 2 31E 06 0 52 Zv F8 727UVA Wnt3a 18575 100 00 4 47E 08 100 00 Zv F8 3237 9048 48 71 3 51E 06 0 79 L F8 11361 61 16 4 31E 06 0 96 2 68 089199 7054 37 98 3 83E 06 0 86 Zv F8 8104 43 63 4 59E 06 1 03 Zv F1 TZTULAT Wnt3as 27 98 1 64E 07 19 74 Zv ki 29 44 8 33E 07 100 00 Zv ki 2201 20 72 1 71 07 20 52 Zv h 28 53 2 83E 07 33 95 Zv ki 18 74 2 36E4 07 28 30 Zv F1 1585367 2074 19 53 2 63E 07 31 55 Zv k1 189568 1432 13 48 1 03E 07 12 39 FL H1 2503 24 42 2 62E 07 31 43 105 2011 526786 2011 10 20 E
110. 07 34 48 FL H4 4 234 ZVL k4 1693136 4881 129 16 3 20E 07 32 73 Zv 4 15833137 1740 46 05 9 16 6 9 35 Zv k4 18430 168 46 4 22E 07 43 13 20 30 40 107 2011 526786 2011 10 20 ERROR RI i 17 89 34185 JL as Zv ka4 204 32 3 07 31 37 Zv ka4 94 56 1 07 18 32 7 103 00 1 12E 07 11 46 Zv k4 161 77 3 45E 07 35 22 Zv F4 1t 141 4310 114 04 1 61E 07 16 48 ZU k4 134 71 3 74E 07 38 22 Zv ka4 174 65 8 50E 07 86 77 Zv k4 94 79 2 17 07 22 14 2 4 181144 6787 179 57 7 55 69 2 ZU k4 67 59 1 83E 07 18 71 210 4 87 03 1 48E 07 15 11 ZU k4 101 06 2 34E 07 23 93 1114 1259 33 32 1 34E 07 13 67 Zv F4 158522 5517 7 07 ZvL k4 1855150 5104 3 34E 07 20 05 Wnt3a 7159 8 12 6 2 5 173 09 3 2 08 100 00 Zv k5 45 89 1 03E 06 0 32 21 05 18152 4693 76 50 3 08E 06 0 95 7 05 158 40 1 20 06 _ 0 37 Zv k5 56 70 1 97E 06 0 61 20 05 152 28 3 45 06 1 07 FU ES 6 68 37 2 52 06 80 6 207 40 3 14E 08 00 JZv h6 1859157 2480 51 94 1 22E 06 0 39 20 66 110 41 1 30 06 0 41 ZL ke 20 66 88 21 1 05 0 04 Zv ke 51 09 9 20 05
111. 1 25uM GDF8 it 34 2 5uM amp 34 SuM 490 490 a GDF8 G K3i BIO 2 5uM GDF8 G K3i BIO GDF amp GSK3i BIO 10uM DIE 2 0 490 OD 490 GOF3 2940 2 5 GDF8 it e 40 Sum GDFS 15 40 10uM 52 GDF3S t 6156 1 25uM GDF8 t615 56 2 5uM GDF8 I amp 56 SuM 150 2011 526786 2011 10 20 120 AA wnt3a GDF8 MCX GDF8 wnt3a Pdx1 ddCT T AA wnt GDF8 MCX CD99 27 03 s 2 5 dx 215 24 Cdx2 1614 1567 965 1 5 m DE HNF4a 339 210 267 1 E 7 23 AA wnt3a GDF8 216 GDF8 wnt3a 600 500 400 7 300 200 100 0 AA wnt GDF8 MCX GDF8 wnt Pdx1 ddCT 57 024 276 545 71 2 1971 2496 983 HNF4a 640 426 609 151 2011 526786 2011 10 20 INTERNATIONAL SEARCH REPORT sii PCT US2009 049183 A CLASSIFICATION OF SUBJECT MATTER C12N5 06 According to International Patent Classification PC or to both national classification and IPC B FIELDS SEAHCHED Minimum documentatlon searched classification system followed by ciassification s
112. 1 37 1 24 7 29 5 18 3 18 8 27 1 223 22 6 17 6 21 3 18 6 224 299 229 22 7 25 5 28 2 32 27 8 25 5 23 23 2 27 8 27 5 30 9 22 8 82 1 ftris GAPDH ALB NEURODI NEUROG3 2 2 2 5 NKX6 1 POU3F4 557 E 18 3 31 3 22 7 24 1 32 5 25 4 29 7 32 28 251 242 26 9 29 37 5 22 5 28 9 19 3 31 1 23 4 246 32 3 25 6 29 9 30 5 28 1 254 246 29 9 299 33 4 221 32 1 21 1 32 1 24 9 248 33 8 26 3 34 30 2 20 7 73 25 9 29 7 31 5 347 221 83 3 tanse METIS RAS GAPDH 2 cpx2 cce ins isti NEURODI NEUROG3 2 5 6 1 Paxa PAX6 Poxi POU3F4 PTFIA 557 2101 asi 20 4 24 8 30 7 276 25 7 19 4 247 20 5 264 34 6 27 2 271 40 29 6 30 7 38 4 9 286 29 2 81 3 32 8 384 222 84 8 20 7 35 29 32 3 30 5 30 7 278 28 5 32 3 31 5 33 9 2241274 21 3 34 5 29 2 31 6 38 1 30 4 28 1 29 32 9 33 9 87 7 22 1 84 5 jou 21 2 34 8 28 1 33 8 30 7 29 3 27 2 27 9 33 6 33 2 352 21 34 9 10 20 127 2011 526786 2011 10 20 00000 14 fare
113. 10 H H DOLO EDEN ES 20 H 30 40 50 62 2011 526786 2011 10 20 C 2 53 A 00000 AL C G C
114. 137 1 4 5 7 w 12 10 vot 2 08 E 5 0 6 amp 04 0 2 m h 10 30 100 300 10 30 100 300 10 30 100 300 TGFb1 ng ml TGFb1 ng ml ng ml 34 13 34 Wnt3a 14 4 12 Sox175878 10 ES M 0 8 0 0 6 3 04 3 02 0 0 a 10 30 100 300 10 30 100 300 10 30 100 300 BMP2 ng ml BMP2 ng ml BMP2 ng ml Wnt3a Ik 2 2 34 34 Wnt3a 300 000 000 250 000 000 200 000 000 ES T 150 000 000 5 2 100 000 000 50 000 000 hi S e E E e 3 3 KS ES S 4 P x 300 000 000 250 000 000 200 000 000 w 150 000 000 5 100 000 000 50 000 000 0 156 0 313 0 625 1 25 25 5 10 AA 588 582 181 p M 2011 526786 A 2011 10 20 DOTEE 100ng mLOAAI 1 4 124 BMP3 ng ml 1 0 0 8 0 6 4 04 0 2 00 10 30 100 300 ng ml ng ml 34 34 Wnt3a DINTIERAL 100ng mLODAA 14 12 7 1 0 0 8 0 6 0 4 02 00 10 30 100 300 10 30 100 300 10 30 100 300 BMP4 ng ml BMPA ng ml BMP4 ng
115. 15 3 9 1 GDF8 24216 22 4 8 75E 06 2 30E 06 26 32 2 5 1 GDF8 2217 19 1 8 752 06 1 94E 06 22 16 2 5 1 GDF8 124218 8 8 6 44E 06 9 70E 05 15 05 9 1 GDF8 18581219 2 5 1 79E 06 1 29E 06 72 17 0 5 TOTULAT 2 Wnt3a8 103 2 2 24 07 0 00E 00 0 00 6 7 132 2011 526786 2011 10 20 Esndeooige i 18 Sox17561H5 Zv k sp cv 9 3E49 685 sp cv 2 Wnt3a 15290 1119 7 32 100 0 3 37E 08 2 84 07 8 44 100 0 2 GDF8 1580202 20177 987 4 89 132 0 4 8 08 1 94E 07 4 00 144 0 2 GDF8 12227 2911 4619 158 69 19 0 3 89E 07 6 69E 07 172 00 11 5 2 GDF8 16215 4383 1775 40 49 28 7 3 57E 07 3 57E 07 100 03 10 6 2 GDF8 911228 4043 1253 30 98 26 4 3 10 07 2 53E 07 81 62 9 2 2 GDF8 1 amp 229 3451 892 25 85 22 6 1 80E 07 1 46E 07 81 07 5 4 2 GDF8 3163 805 25 44 20 7 1 58E 07 3 54E 06 22 32 4 7 GDF8 166220 2791 1453 52 05 18 3 1 40 07 9 06 64 28 4 2 2 GDF8 198815 3137 1172 37 34 20 5 1 30E 07 7 52E 06 57 85 3 9 2 GDF8 169230 2624 248 9 46 17 2 1 24E 07 1 55E 07 124 73 3 7 2 GDF8 15814231 31 2 1 22 07 6 51E 06 53 37 3 6 2 GDF8 65232 21 4 1 18E 07 1 51E 07 127 98 3 5 2 GDF8 12221 12 8 1 18 07 1 54E 07 181 11 3 5 2 GDF8 1
116. 20 SE l 30 40 50 C 2011 526786 2011 10 20 88 l 10 20 30 40 50
117. 24 April 2008 2008 04 24 Claims 116 123 paragraphs 0173 0260 0293 ARMSTRONG LYLE AL The role of MAPK ERK and NF kappa beta signalling in the maintenance of human embryonic stem 1 pluripotency and viability highlighted by transcriptional profiling and functional analysis HUMAN MOLECULAR GENETICS OXFORD UNIVERSITY PRESS SURREY vol 15 no 11 1 June 2006 2006 06 01 pages 1894 1913 002471543 155 0964 6906 the whole document PALING N R D ET AL REGULATION OF EMBRYONIC STEM CELL SELF RENEWAL BY PHOSPHOINOSITIDE 3 KINASE DEPENDENT SIGNALING JOURNAL OF BIOLOGICAL CHEMISTRY AMERICAN SOCIETY OF BIOLOCHEMICAL BIOLOGISTS BIRMINGHAM US vol 279 no 46 24 August 2004 2004 08 24 pages 48063 48070 XP002380573 ISSN 0021 9258 the whole document HORI YUICHI ET AL Growth inhibitors promote differentiation of insulin producing tissue from embryonic stem cells PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF USA NATIONAL ACADEMY OF SCIENCE WASHINGTON DC US vol 99 no 25 10 December 2002 2002 12 10 pages 16105 16110 XP002392968 ISSN 0027 8424 cited in the application the whole document MCKIERNAN EADAOIN ET AL Directed differentiation of mouse embryonic stem cells into pancreatic like or neuronal and glial like phenotypes TISSUE ENGINEERING OCT 2007 vol 13 n
118. 28 6 28 2 25 4 27 0 24 2 26 9 26 2 32 0 24 4 27 2 312 28 0 80 1 25 8 85 1 28 8 292 315 28 2 324 23 1 24 0 2202 18 6 19 2 280 22 6 234 149 213 15 0 227 31 8 22 6 21 9 28 6 23 7 29 9 25 8 26 9 24 7 236 277 292 27 9 19 4 32 8 AA 483940 18 3 18 9 27 9 22 3 23 0 14 6 21 1 14 7 22 5 31 5 22 4 21 6 29 0 23 5 32 9 26 1 27 1 24 4 232 281 292 28 1 19 1 31 9 AMEES 18 3 13 1 28 0 23 0 226 15 1 20 5 15 1 22 8 31 8 22 8 22 1 29 5 24 2 33 8 Be 27 4 24 9 238 28 3 9 29 7 19 5 32 0 AA 12206 18 2 19 5 279 222 23 4 14 4 21 3 14 8 22 5 31 1 224 21 7 280 23 6 35 8 25 9 27 1 24 1 233 290 28 7 27 5 19 7 32 7 GDF8 174 205 8 2 25 2 24 4 181 228 17 7 3 8 233 24 9 1 25 9 31 4 26 6 294 266 256 297 27 8 296 211 225 GDF8 Wnt3a 17 8 20 5 28 2 24 8 24 3 17 7 22 9 17 5 24 2 31 9 23 5 24 0 30 2 25 6 31 9 27 0 29 3 26 5 256 298 28 1 30 7 21 5 22 8 GDF8 181 18 0 19 1 27 6 224 23 4 14 5 21 2 14 8 22 5 31 5 22 6 212 274 234 33 8 25 0 26 8 24 4 23 0 27 5 29 8 27 3 19 5 31 9 GDF84 62180 18 0 18 0 27 3 22 2 229 142 209 14 4 22 2 31 4 22 1 21 2 279 23 2 40 0 25 1 26 1 238 229 296 28 7 27 3 18 8 31 5 GDF8 18 3 18 5 27 8 234 23 0 16 2 21 2 15 6 232 32 2 23 2 22 6 312 24 3 33 9 28 6 27 3 25 2
119. 3 000 2 000 2 000 1 000 1 000 0 0 0 078 015 0 43 0625 125 25 10 0 0 078 0 156 03138 0625 125 25 5 10 GDF8 3818 229 19 u 3819 40 u M 3818 6 CxCR4 5 000 10 4 000 Y a 4 3 000 50 2 000 1 000 EI g gt Si g P 0 0 078 0 155 0313 0625 125 25 10 S amp GDF8 34 u M g 6 000 12 5 17 5 000 20 8 4 000 06 504 ES i E 3009 0 2 0 2 000 D p gt S S ES E 1 000 lt A 2 59 lt 8 g 0 0 0 078 0 156 0 313 0 625 125 25 GDF8 GSK3i u M 3553 2011 526786 2011 10 20 145 Wl INS 1 5 usu Z YY e YY 0 0 ins GcGo Et s 1 5 Em 2 50 17 1 e 5 THOXO ESE 21 6 212 22 2 214 223 22 6 223 23 4 233 25 1 229 25 6 256 25 7 25 5 258 244 20 4 220 22 0 220 177 220 227 22 2 19 6 CXCR4 F
120. 36 3 33 7 40 0 GSK3 18 4 249 30 3 31 5 222 347 21 7 34 5 29 9 35 3 27 6 29 2 26 8 27 9 34 6 33 9 30 6 35 1 240 30 0 23 6 40 0 GDF8 206 18 2 27 6 30 2 33 5 23 8 40 0 22 9 40 0 27 9 35 1 27 1 29 9 27 7 29 3 40 0 32 3 31 3 27 4 233 32 7 30 8 330 29 2 30 3 RT PCR GAPDH ALB ARX INS 1811 MAFB NEUROD1 NEUROGS 2 2 NKx2 5 1 paxo Poxi 4 sst 2101 19 0 224 28 6 23 9 23 7 22 8 22 1 23 6 24 1 30 6 23 5 24 7 26 8 24 0 31 9 27 2 25 7 27 1 23 7 29 8 26 5 29 6 243 32 5 wnt3a 19 5 23 5 29 3 24 6 24 1 25 1 22 7 25 2 25 1 31 1 24 4 24 4 24 1 24 5 31 8 27 5 26 1 27 9 241 27 2 91 1 25 9 32 4 AAT A181 18 0 210 27 7 240 22 1 24 5 210 24 6 24 5 31 6 24 0 24 2 24 6 24 6 30 9 28 1 26 28 0 240 279 275 31 0 25 5 31 2 AAT 12180 194 210 27 3 26 3 216 254 206 267 26 1 347 4 6 255 25 8 25 0 29 4 289 977 28 9 240 567 285 32 6 27 1 312 AA tano RNASEZZ VIRES AAT ite 1202 19 2 20 7 29 1 24 0 233 21 8 219 22 7 244 a1 1 239 240 24 1 24 4 32 1 27 2 26 3 26 0 243 296 277 29 5 24 0 35 1 AAT 154040 19 2 20 8 29 4 24 7 22 9
121. 66 1 6 3 GDF8 1221 2 3 2 36E 06 3 07 06 129 78 0 7 FIFEVATE 4 Wnt3a EL 119 3 2 42 07 00E 00 00 7 8 BH D Wu 10 20 30 134 2011 526786 2011 10 20 0000 00 18 7 Zv k 2185 FIORITO FARRE sp CV 4 T2TUVA Wnt3a 13945 100 0 3 09E 08 4 77E 07 15 46 100 0 4 GDF8 1534 7416 53 2 2 10 8 1 82E 08 86 70 68 0 4 GDF8 1549240 11283 80 9 1 61E 08 4 41 07 27 34 52 2 4 GDF8 13428 5236 37 5 4 07 3 08E 07 76 36 13 1 4 GDF8 62198 3985 28 6 3 89E 07 5 55E 07 142 91 12 6 4 GDF8 54196 4861 34 9 3 03E 07 1 98E 07 65 37 9 8 4 GDF8 1 1 1921 13 8 2 94E 07 3 65E 07 123 90 9 5 4 GDF8 124186 3486 25 0 2 34E 07 1 42 07 60 78 7 6 4 GDF8 181254 3960 28 4 2 31 07 2 27 07 98 10 7 5 4 GDF8 24168 3460 24 8 2 28 07 7 13 06 31 23 7 4 4 GDFS 1580190 3402 24 4 1 87E 07 1 58E 07 84 61 6 1 4 GDF8 162 1255 28 7 1 52 07 1 07 68 91 4 9 4 GDF8 1 250 19 1 1 48E 07 8 30E 06 56 15 4 8 4 GDF8 08127 26 7 1 19 07 9 69E 06 81 29 3 9 4 GDF8 10184256 21 0 9 41 06 8 6 06 92 01 3 0 4 GDF8 144257 22 8 8 06E 06 4 49E 06 75 2 6 4 GDF8 15258 19 6 7 89E 06 7 24E 06 91 70 2 6 4 GDF8 15
122. 7485 47 4 8 10 07 28 3 it amp tna 1589017 1 13E 08 45 0 3416 9 20E 07 36 7 8318 1 01E 08 35 4 taga tams TE 1000 10 20 30 40 121 2011 526786 2011 10 20 Ho BE HI E 7 Zv HER paemes penge 1 9164 149 46 5 91 06 5 17 1 10ng mLODAA ZU 6132 100 00 1 52E 06 1 38 1 100 aL 9658 157 51 1 14E 08 100 00 EGF FGF4 PDGF A VEGF PDGF D 1 10ng mLODAA 988122 ALE JuU GDF8 Wnt3a 8 78E 07 76 82 1 10ng MLMAA 2122 EGF FGF4 PDGF AB VEGF Wnt3a 4 70E 07 41 09 1 10ng mLDAA 2122 EGF FGF4 PDGF A 4 E J Wnt3a 4 42E 07 38 65 1 10ng MLQAA 184122 EGF FGF4 PDGF AB Wnt3a 3 43 07 30 03 1 10ng mMLQAA 10488122 EGF FGF4 Wnt3a 4 59 07 40 13 1 1O0ng mLOAA ite 922 FGF4 Wnt3a 4 28E 07 37 43 1 10ng mLOAA 184192 EGF Wnt3a 2 80E 07 24 45 1 10ng mMLQAA 181122 Wnt3a 3 7 26 65 EGF FGF4 PDGF A VEGF PDGF D 1 10ng mLDAA 1 2 35 A2E Ju GDF8 Wnt3a 4 86E 06 4 25 1 10ng mLDAA 162035 EGF FGF4 PDGF AB VEGF Wnt3a 96E 06 46 1 1535 EGF FGF4 PDGF A 4 E JL Wnt3a 1 672 06 1 46 1 10ng mLQAA 1235 EGF FGF4 PDGF AB Wnt3a 4 56E 06 3 99 1 10ng mMLQAA 835 EGF FGF4 Wnt3a 4
123. EGF 100ng mL 16336 128 87 2 32E 08 85 24 984194 Wnt3a amp 15 FGF 100ng mL 19853 156 61 2 59E 08 95 45 Wnt3a amp 4 FGF 100ng mL EGF 25ng mL 19880 156 83 2 59E 08 95 47 16834 Wnt3a FGF 100ng mL EGF 50ng mL 18166 143 30 2 35E 08 86 30 088194 Wnt3a 100 mL EGF 100ng mL 11241 88 68 1 55E 08 57 10 EL Wnt3a amp EL EGF 50ng mL 5558 43 85 5 1 07 18 44 Ab Wnt3a amp 8 EGF 100ng mL 6818 53 79 6 42 07 23 62 Wnt3a 5 FGF 50ng mL 8494 67 01 6 62 07 24 35 AAT E Wnt3a t FGF 50ng mL EGF 50ng mL 10138 79 98 7 07 26 87 AL AAT E Wnt3a t FGF 50ng mL EGF 100ng mL 10219 80 62 7 75 07 28 51 E Wnt3a amp t FGF 100ng mL 9944 78 45 6 68E 07 24 59 3 Wnt3a FGF 100ng mL EGF 50ng mL 11046 87 14 8 17 07 30 07 AAT E Wnt3a8 t FGF 100ng mL EGF 100ng mL 7695 60 71 6 87E 07 25 28 3811 797 GDF8 sez 1600 10000 9 20 10000 9 00 soo 7 10000 660 8490 6 30 400 200 100 50 25 12 50 soso 6 20 3 13 3 1 90 0 10 20 56 11 020 ozo oso 10 20 30 40 12 Y h 4 AFP cp 9 cp9 2 CXCR4 Hs0023705
124. amp EGF 6145 85 53 1 5E 07 19 18 Wnt3a amp FGF4 5323 74 09 2 7E 07 33 75 1834 Wnt3a 8 PDGF D 5017 69 84 1 07 18 76 8194 AAT Wnt3a amp 8 PDGF A 4175 58 11 1 1E 07 13 43 1634 AAT Wnt3a 2 f VEGF 4713 65 60 1 0E 07 12 49 62934 AAT Wnt3a 2 GDF8 6354 88 44 7 1E 07 88 59 AAF LIETI 7286 101 41 3 1E 07 38 38 1683384 Wnt3a amp A PDGF D VEGF 5030 70 01 1 2 07 14 58 62934 AAT VEGF ALE 776 10 81 1 06 1 56 AAFS Wnt3a g PDGF D LVE JL 3490 48 57 6 5E 06 8 02 083194 WntSa amp GDF8 PDGF D 6889 95 88 5 8E 07 72 59 AATA Wnt3a PDGF D AL E JL VEGF 2133 29 68 2 7 06 3 32 AATA Wnt3a GDF8 PDGF D VEGF 5585 77 74 6 6E 07 81 75 1089194 AAT 3 Wnt3a gA GDF8 VEGF 9 6083 84 67 5 6E 07 69 62 156 34 AAT GDF8 PDGF D VEGF AL E L 9455 131 60 9 6E 07 119 24 15434 AAT Wnt3a EGF FGF4 PDGF A VEGF PDGF D AY E JL GDF8 4757 66 21 3 9E 07 48 77 1189194 AAT Wnt3a amp EGF FGF4 PDGF A VEGF PDGF D AZE JL GDF8 6028 83 90 7 0E 07 87 44 10 20 30 40 50 116 2011 526786 2011 10 20 a Sox173838 mg Fi 165189 1151896 FIFEVA TRE Wnt3
125. ml Wnt3a tt amp 34 15134 Wnt3a SOX1738PE 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 9 9 078 0 156 AA 381 515180 1870 0 313 0 625 1 25 25 5 10 Sox1 755 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 9 0 078 0 156 0 313 AA 38 tt amp t219 u M 188 0 625 1 25 25 5 10 138 2011 526786 2011 10 20 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 4 0 078 0 156 0313 0 625 125 AA 34 M 188 ZA 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 9 0 078 0 156 0 313 0 625 1 25 25 AA GSK3i BIO y M 1818 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 0 078 0 156 0 313 0 625 125 AA 3818 1283 19 u M 36 25 300 000 000 250 000 000 200 000 000 el el T 150 000 000 E x x 5 5 100 000 000 50 000 000 0 9 0 078 0 156 0 313 0 625 1 25 25 5 10 3818 683 202 u 1 300 000 000 250 000 000 200 000 000 150 000 000 E x x o o
126. 0 30 40 50 2011 526786 2011 10 20 11 10 20 30 l
127. 0 29 2 6 1585162 1260 26 39 1 35 06 0 43 7 20 07 328 17 4 92E 08 100 Zv rF7 270 81 7 31E 06 1 49 108 2011 526786 2011 10 20 10 10 00 EREDE RI we T J 20 07 462 165 3015 49 32 7 34E 06 1 49 20 07 155835166 13845 226 45 7 98E 06 1 62 Zv k7 168 87 7 35 6 1 49 Zv k7 231 26 6 49E 06 1 32 Zv k7 78 08 6 38E 06 1 30 2 FIFELARE Wnt3a fA 6868 179 83 2 31E 06 0 52 8 486 35 4 47E 08 100 00 8 344 04 4 1 06 0 93 Zv ka Zv te 8 307 91 3 33E 06 0 75 Zv F8 62175 5 0 13 4 91E 06 1 10 Zv F8 15893177 9994 261 68 2 95E 06 0 66 FL H8 285 58 3 74 06 0 84 2 5 TZTULAT Wnt3a amp d 7159 67 42 8 12E 06 2 51 2 5 100 00 3 23E 08 100 00 Zv ks 44 44 1 96E 06 0 61 20 05 2036 19 18 1 79E 06 0 55 FL ES 24 13 1 56E 06 0 48 20 65 42 09 7 05 05 0 22 FL H5 57 30 3 08E 06 95 20 05 26 7276 68 52 2 38E 06 0 74 2 5 13427 4588 5 63E 05 2 5 03128 2682 1 37E 06 2 5 158929 5778 1 94E 06 FU ES 70 05 27 66 PIFELARE 69 07 2 52E 06 O 80 2 6 FIFEVA Wnt3a 9902 209 51 3 14E 08 100 00 FLH6 45 32 1 33E 06 0 42 20 66 e
128. 0 9 36 0 25 8 25 0 23 2 22 2 25 2 23 7 28 5 249 228 34 3 23 4 32 7 83 5 AAT 18 206 19 8 40 0 24 9 2 7128 L 21 7 40 0 18 3 19 6 20 7 96 2 24 4 24 8 26 7 22 6 24 1 4008 23 9 220 33 2 23 3 32 4 34 6 GDF8 21 6 40 0 25 5 25 9 25 2 22 3 40 0 20 1 24 6 22 1 34 9 25 9 27 5 24 8 24 2 25 3 25 4 30 6 250 243 29 5 25 2 29 8 82 9 GDF8 Wns3a 212 aoo 250 25 251 226 400 197 24 6 220 25 7 27 8 24 4 23 8 250 25 1 305 248 244 81 1 25 0 45 32 6 181 20 7 40 0 25 1 23 6 25 5 22 4 40 0 20 0 23 6 21 5 36 3 25 2 25 4 23 9 23 6 24 3 26 3 28 1 25 4 23 3 31 8 23 7 32 8 31 9 GDF8 tamiso 20 9 400 25 6 24 0 26 9 22 1 347 19 9 23 0 21 1 36 6 25 0 25 2 23 9 23 6 24 2 25 8 28 3 26 3 230 33 2 24 2 32 8 32 3 GDF8 9119 196 40 0 28 9 23 7 24 6 20 7 40 0 18 0 22 7 20 4 33 2 24 2 25 0 22 8 22 1 24 0 22 6 28 5 23 1 22 7 30 3 22 6 32 1 29 3 GDF8 169202 18 5 306 22 1 20 2 22 8 19 7 35 8 18 5 21 8 19 9 34 3 23 6 22 7 92 5 23 5 23 1 25 9 27 7 251 21 5 30 0 22 3 32 0 32 5 GDF8 83409 197 400 23 0 226 24 7 20 5 33 8 18 2 22 4 20 5 35 4 24 9 24 2 LIRA 23 4 23 4 25 4 28 5 24 0 22 8 31 8 23 7 31 2 32 7 GDF8 GSK3 AMEESBIO 19 6 23 1 21 8 24 3 20 0 334 17 7 233 20 1 34 8 24 7 24 5 22 4 22 7 24 1 25 0 29 3 248 218 31 7 23 3 33 7 84 6 2206 19 7
129. 00 4 000 3 000 2 000 1 000 0 0 078 0 156 0 313 0 625 125 25 5 10 GDF8 388 145 180 u 1811 6 000 5 000 4 000 3 000 2 000 1 000 5 0 0 078 0 156 0 313 0 625 125 25 5 10 GDF8 38 202 1815 2 6 000 5 000 4 000 3 000 2 000 1 000 0 0 078 0 156 0 313 0 625 125 2 5 5 10 GDF8 15832 40 M 1858 6 000 5 000 4 000 3 000 2 000 1 000 5 0 0 078 0 156 0 313 0 625 1 25 25 5 19 GSK3i BIO 1888 2 6 000 5 000 4 000 3 000 2 000 1 000 0 0 078 0 156 0 313 0 625 1 25 25 5 10 GDF8 62 34 p M 1879 6 000 5 000 4 000 3 000 2 000 1 000 0 0 0 078 0 156 0 313 0 625 1 25 25 5 10 GDF8 181 u M SARI 144 2011 526786 2011 10 20 6 000 6 000 5 000 5 000 4 900 4 900 8 m 3 000 3 000 2 000 2 000 1 000 1 000 0 0 078 0 1556 0 43 0625 125 25 10 0 0 078 0486 0 819 0625 125 25 5 10 GDF8 5819 180 M 5813 GDF8 3810 tt amp 202 u M 3818 6 000 6 000 5 000 5 000 4 000 4 000 ES ES m 3000 m
130. 000 00000000 100 00000 00000000000 10000000000000000000 30 wp AVO DR EE 40 37 2011 526786 2011 10 20 2 0 NH NH 7 7 0 000000 00000 1000000 00000 0000 0 0000000000 00000 000 1000000000000 2 mS NH 0 0 c atta 1000000000 00000000 1000000000 00000000 1000 0000 00 000000 Ma a as EISE L C N N Na N d N 10000000 000 1000000000000 0000 1000000000000000 100000000000000000A
131. 00000 10000000000000000000 Percent of Sox17 4 3 4500 05 10 15 20 25 30 35 40 Log ng ml 2011 526786 2011 10 20 2 10 20 n definiti ve endoderm i neage 30 l n 40 l 100
132. 144 37 1 45 07 8 48 77 3117 34 86 1 41 06 0 72 72 8942 100 1 95E 08 100 00 158 135 AATA WntSa amp EGF 19334 216 28 6 62E 07 33 86 1093195 AATA amp PDGF AB 16662 186 34 4 95E 07 25 33 1093135 amp 16885 188 84 4 48E 07 22 94 1835 AAT Wnt3ag VEGF 18263 204 25 3 51E 07 17 98 18135 AAT Wnt3ag f FGF4 4410 49 32 3 07 17 04 9435 LETI 18867 211 00 2 61E 07 13 35 35 AAT Wnt3a f PDGF C 16642 186 12 1 85E 07 9 46 158 035 AAT Wnt3a A PDGF D 17618 197 1 84E 07 9 41 109395 AATA Wnt3a fA PDGF B 14168 158 46 1 52 07 7 76 ite 35 AAT f PD98059 18877 211 11 1 30E 07 6 64 ite 935 3 Wnt3a BMP1 18849 210 81 1 29E 07 6 59 15135 AAT WntSad LY294002 18374 205 49 1 7 5 28 162935 Wnt3a f BMP4 16748 187 31 8 97 06 4 59 1835 AAT Wnt3ag BMP2 16218 181 38 8 89E 06 4 55 JUD RA 8 1 7515 LEHES ERRARE 131896 4083135 Wnt3agB 7 20111 224 91 8 05 06 4 12 1031958 AAT Wnt3a 0124 16539 184 97 7 54 06 3 86 18435 Wnt3a 8 BMP6 17838 199 50 7 32E 06 3 75 12935 AAT Wnt3a BMP2 7 12042 134 67 7 6 3 62 2935 AAT Wnt3a amp B E2207 19312 215 98 1 95E 06 1 00 158 35 AA
133. 1E 07 19 3 2 PDGF A VEGF PDGF D AY E L GDF8 1581272 6060 38 3 5 46E 07 19 1 2 FGF PDGF A VEGF PDGF D AT TE L GDF8 273 5545 35 1 3 99E 07 14 0 2 EGF FGF PDGF A VEGF PDGF D L EJL GDF8 1589074 10898 68 9 3 91 07 13 7 2 PDGF A PDGF D AZE JL GDF8 083175 4117 26 0 3 01E 07 10 5 2 PDGF A PDGF D AZ E L GDF8 08176 3825 24 2 2 74E 07 9 6 2 FGF PDGF A VEGF PDGF D A1 E JL GDF8 1589577 5928 37 5 2 44E 07 5 2 FGF PDGF A VEGF PDGF D AJ GDF8 158 078 3303 20 9 2 7 7 1 2 FGF PDGF A VEGF PDGF D Av EJ GDF8 448879 4767 80 2 1 85E 07 6 5 2 FGF PDGF A VEGF PDGF D EJ GDF8 11580 2194 13 9 1 22E 07 4 3 2 EGF FGF PDGF A VEGF PDGF D 5572 GDF8 152081 2920 18 5 9 16E 05 0 3 2 teens 1819 11 6 1 7 37 2 EGF FGF PDGF A VEGF PDGF D 4 GDF8 08488 2153 13 6 1 19E 07 4 2 2 FGF PDGF A VEGF PDGF D EJ GDF8 84184 58 0 4 2 94E 07 10 3 2 PDGF A PDGF D 5 GDF8 081856 57 0 4 3 07 10 6 1 T2TULAT U nai A EL 6049 45 2 1 31E 07 5 2 000 0 117 2011 526786 2011 10 20 ERRORS RI
134. 2_m1 FGF17 FOXA2 GAPDH GATA4 GATAG esc MIXL1 Hs00430824 21 NANOG Hs02387400 gl OTX2 Hs00222238 POUSFI 50 17 Hs00610080_m1 ALB Hs00609411_m1 124 2011 526786 A 2011 10 20 10 20 30 ERRORES 12 Y 5 4 ARX Hs00292465 mi 2 GAPDH INS Hs00355773_m1 1511 NEUROG3 Hs00360700 gl 2 2 2 5 NKX6 1 PAX4 PAX6 PDX 1 POUSFA 557 10000 125 2011 526786 2011 10 20 10 20 30 00000 126 2011 526786 2011 10 20 Wnt3a GDF8 13 CTh GAPDH 0099 FGF4 FoxA2 GATA4 GATAS esc MixL1 5 sox17 sox7 T s 19 5 34 7 238 24 1 24 5 21 5 86 8 18 4 227 20 3 5 247 23 7 22 1 22 3 25 23 4 28 238 22 6 31 4 23 5 32 2 323 EA 18 7 36 1 23 23 5 23 3 21 36 2 178 21 9 19 9 33 1 23 8 29 7 21 9 22 1 23 0 23 1 28 6 23 21 9 296 234 31 9 32 2 ARS 18 5 33 23 23 1 33 6 20 8 35 3 17 9 21 3 19 7 32 6 24 23 2 21 7 21 9 24 23 27 5 23 2 21 5 30 1 23 2 31 7 321
135. 4 15903 84 92 3 53E 06 1 90 15845102 AAT Wnt3a amp EGF FGF4 12943 69 11 2 85E 05 0 15 797 Wnt3a A 23253 124 16 1 97E 07 10 59 81 FIFECVA WntSa 18728 100 00 1 86E 08 100 00 91135 8 Wnt3a amp EGF FGF4 18294 97 68 1 99E 07 10 70 1006060 IU O 689339 91 28 15 19 1852337 83 32 14 37 5 10 20 30 40 LED SEE SHE 112 JP 2011 526786 A 2011 10 20 RA nm MI 1 758535 96 ZU 7107 67 96 1 27 07 7 94 FTIFEYA Wnt3a 10459 100 00 1 60E 08 100 00 158 17 Wnt3a amp 6942 73 43 1 27E 06 0 74 158 17 AAT 1 f EGF FGF4 5738 60 69 3 14E 06 1 83 817 Wnt3a amp EGF FGF4 PDGF AB 4453 47 10 5 0 54 2917 AAT 8 Wnt3a f EGF FGF4 PDGF AB 4 E JL 10391 109 91 8 92E 06 5 20 465917 Wnt3a amp i EGF PDGF A VEGF 5728 60 59 2 14 06 1 24 AAT B FGF4 PDGF A VEGF 13198 139 59 1 29 07 7 54 2917 AAT 3 Wnt3a amp EGF FGF4 PDGF A VEGF 10480 110 85 8 97E 06 5 23 1585017 AATA Wnt3a amp EGF FGF4 PDGF A A 7E V 13649
136. 400 22 7 225 232 21 0 29 9 18 4 23 0 20 3 34 5 25 8 249 22 9 23 6 238 25 7 29 7 247 222 30 7 24 4 32 83 8 RT PCR mi GAPDH ALB AMY2A ARX INS isti MAFA MAFB NEUROD1 NEUROG3 2 2 2 5 NKx6 1 PAx4 PAxe PDX1 PousF2 1 SST 201 AA 18 5 269 30 6 32 0 22 9 34 1 22 4 34 8 29 3 36 4 27 4 30 1 280 29 3 34 6 322 31 4 400 233 30 5 346 33 2 34 8 Wnt3a 18 4 27 3 30 2 33 o 23 0 34 7 22 4 40 0 28 8 33 9 27 7 30 4 281 29 0 40 0 349 1 0 38 1 238 308 30 7 40 0 32 8 40 0 AAT tets 18 6 26 0 90 1 34 2 22 3 40 9 22 3 40 0 30 4 34 6 28 5 31 4 29 0 30 3 33 9 35 8 33 3 40 0 250 30 8 36 0 34 6 34 8 AA 2180 18 8 255 300 333 225 35 1 22 5 40 0 294 34 3 286 32 6 31 1 32 5 35 3 35 3 36 2 263 289 31 3 40 0 347 34 4 kemio 34 1 400 40 0 40 0 40 0 40 0 37 4 40 0 40 0 40 0 34 9 40 0 400 40 0 40 0 40 0 40 0 40 0 40 0 400 40 0 40 0 40 0 40 0 42202 18 5 26 2 30 7 33 0 225 34 8 22 6 35 1 29 8 35 1 28 2 30 0 28 5 29 5 53 6 339 32 o 36 6 24 1 29 7 31 9 40 0 34 2 40 0
137. 5 5 3 54E 07 12 4 2 EGF FGF PDGF A VEGF PDGF D GDF8 4158 26 3 8 23 07 11 3 2 FGF PDGF A VEGF PDGF D A2E JL 62194 3470 22 0 2 96E 07 10 4 2 FGF PDGF A VEGF PDGF D A 3E JL GDF8 62195 3800 24 0 2 7 10 3 2 EGF FGF PDGF A PDGF D A2 E JL GDF8 62196 4619 29 2 2 78E 07 27 2 4011 25 4 2 45E 07 8 6 2 tetos 4367 27 6 1 92E 07 6 7 2 ireto 3162 20 0 1 20E 07 4 2 2 EGF FGF PDGF A VEGF PDGF D GDF8 15832200 2087 13 2 4 06 1 6 2 EGF PDGF A VEGF PDGF D 3 2 GDF8 158 0201 1568 9 9 6 17 06 2 2 2 EGF PDGF A PDGF D A1 JL GDF8 15202 5213 33 0 1 41 07 4 9 2 PDGF A VEGF PDGF D GDF8 1981203 7 0 0 3 04E 07 10 6 2 EGF FGF PDGF A VEGF PDGF D AL E JL GDF8 204 11 0 1 3 18 07 11 1 2 EGF FGF PDGF A VEGF PDGF D AY E L GDF8 1 205 10 0 1 3 20 07 11 2 10 20 30 120 2011 526786 2011 10 20 35 ARX 80 17 9 kanas visiere TE tamo Um 1688321 1 65E 08 65 9 65 913 1 612 08 64 3 1 035 1 59E 08 55 7 tame m 1333
138. 50 000 000 9 0 078 0 156 0 313 0 625 125 25 5 10 GDF8 359 181 1218 17 Sox1738f 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 0 0 078 0156 0 313 0 625 125 2 5 5 10 GDF8 152 180 187 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 9 0 078 0 156 0 313 0 625 1 25 25 5 10 GDF8 SAM 202 u M 1518 Sox1 1 7586 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 9 0 078 0 156 0 313 0 625 GDF8 162 19 u M 1878 1 25 25 5 10 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 9 0 078 0 156 0 313 0 625 1 25 25 5 10 GDF8 358 40 140 2011 526786 2011 10 20 Sox1 7585 1 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 9 0 078 0 156 0 313 0 625 1 25 25 5 10 GDF8 3 815 34 y 1 Sox1 7551 300 000 000 250 000 000 200 000 000 150 000 000 100 000 000 50 000 000 9 0 078 0 156 0 313 0 625 125 25 5 10 GDF8 388 25 181 u sara 7
139. 53E 08 4 08E 07 26 72 43 1 3 GDF8 40831358 48 8 1 35E 08 4 86 07 36 08 38 0 3 GDF8 1 amp 22 42 4 1 06E 08 04E 07 47 73 29 8 3 GDF8 30 8 4 31E 07 2 11E 07 48 95 12 2 3 GDF8 1181184 26 3 3 94E 07 2 24E 07 56 78 11 1 3 GDF8 1 2 amp 223 26 7 3 55E 07 2 51 07 70 56 10 0 3 GDF8 185237 26 4 3 11 07 1 56E 07 50 18 8 8 3 GDF8 1 244 28 2 2 7 7 1 52 07 55 71 7 7 3 GDF8 245 29 8 2 59 07 1 03 07 39 90 7 3 3 GDF8 115246 24 1 2 0 07 2 69E 07 117 08 6 5 3 GDF8 1 247 23 5 2 14 07 1 04E 07 48 63 6 1 3 GDF8 154248 24 4 2 13E 07 9 45E 06 44 45 6 0 3 GDF8 1 825 24 4 1 76 07 1 21E 07 69 04 50 3 GDF8 483195 23 3 1 61E 07 27 06 20 34 4 5 3 GDF8 129227 18 3 1 07 1 07 94 25 4 0 3 GDF8 1699183 3306 969 29 32 20 8 1 35 07 1 14 07 84 25 3 8 3 GDF8 3187 17 4 1 35 07 9 02E 06 66 67 3 8 3 GDF8 62201 20 3 1 35E 07 1 69E 07 125 02 3 8 3 GDF8 81197 20 6 1 7 5 2 06 40 51 3 7 3 GDF8 1971249 24 2 1 29 07 6 79E 06 52 72 3 6 3 GDF8 168141 15 1 1 12 07 4 95E 06 44 30 3 2 3 GDF8 24194 20 0 9 75 06 2 11E 06 21 63 2 8 3 GDF8 129250 23 2 9 14E 06 4 78E 06 52 32 2 6 3 GDF8 15251 19 0 8 412 06 4 59E 06 54 60 2 4 3 GDF8 17 6 6 77 06 1 86 06 27 45 1 9 3 GDF8 159252 16 3 6 20 06 2 1 05 3 72 1 8 3 GDF8 15253 15 7 5 83E 06 1 47E 06 25 20 1 6 3 GDF8 15814202 13 2 5 752 06 2 62 06 45
140. 580233 19 9 1 12E 07 1 07 97 44 3 3 2 GDF8 24147 22 5 1 12 07 9 80E 06 87 75 3 3 2 GDF8 2234 18 5 9 47E 06 5 67E 06 59 84 2 8 2 GDF8 5235 22 2 9 10 06 6 51E 06 71 52 2 7 2 GDF8 166236 18 8 6 73 06 6 32E 06 93 82 2 0 2 GDF8 403199 15 4 6 60E 06 2 45E 06 37 20 20 2 GDF8 iAH 21 0 6 48E 06 3 09E 06 47 67 1 9 2 GDF8 158 53 18 2 6 36E 06 2 89E 06 45 36 1 9 2 GDF8 1855237 19 4 6 02 06 3 00E 06 49 79 1 8 2 GDF8 1520238 17 9 5 97E 06 6 10 06 102 07 1 8 2 GDF8 29239 20 6 5 60E 06 2 42E 06 43 24 1 7 2 GDF8 491240 19 6 4 68E 06 8 13E 06 66 80 1 4 2 GDF8 1081200 15 21 4 39E 06 1 88E 06 42 83 1 3 2 GDF8 484222 11 6 3 33E 06 2 52E 06 75 78 1 0 2 18241 19 3 3 29E 06 1 55E 06 47 03 10 2 GDFS Aire p242 2 5 1 08E 06 8 85 05 81 61 0 3 2 GDF8 1580243 1 6 2 53E 05 3 07E 05 121 25 0 1 2 GDF8 5204 1 6 1 36E 05 2 27E 04 16 66 0 0 3 Wnt3a amp EL 99 6 2 82 07 00E 00 00 8 0 HH D BW 133 2011 526786 2011 10 20 ERRORS 18 Sox17 H5 Zv k LES FAA SD cv AEREA SD cv 3 FIF ZA Wnt3a 15867 785 4 95 100 0 3 54E 08 2 40 07 6 77 100 0 3 GDF8 6974 3723 53 38 44 0 2 07 08 9 51 07 45 85 58 6 3 GDF8 184185 10892 1552 14 24 68 6 1
141. 8 1 48E 06 ZL H8 4 44 06 2 46 2 85 3 48E 06 2 68 1 86 310 54 4 22 06 0 94 8 231 58 4 07 06 91 Zv k8 1688188 10419 448 27 4 27 06 0 96 506 83 3 94 406 0 88 ZL ka 20 68 1589291 6224 267 78 4 53E 06 1 110 2 Sox1758JE 138 887 99144 589785 1589130 105 104 i620 iL amp 22 62 37 129 3 29 1 1629134 126 9 35 0 29150 142 it amp t136 9580 102 2089 L amp 105 TE 133 000 2011 526786 2011 10 20 10 20 30 111 2011 526786 2011 10 20 3 SRO 7 pus 4185 FIORA fe EL FIFCVAT Wnt3a8 23253 124 16 1 97E 07 10 59 84 FIFCVA Wnt3a 18728 100 00 1 86E 08 100 00 1585517 AAT Wnt3ae EGF FGF4 21445 114 51 3 4 07 18 48 EL FIFCVAT Wnt3a 23253 124 16 1 97E 07 10 59 81 Wnt3a 18728 100 00 1 86E 08 100 00 22 AA Wnt3a f A EGF FGF4 18336 97 91 3 72E 07 20 05 AATA WntSa f EGF FGF4 18891 100 87 3 26E 07 17 55 0890129 AAT E
142. 8 189 16 9 7 72 06 5 36E 06 69 41 2 5 4 GDF8 115259 15 7 6 92E 06 2 58E 06 37 29 2 2 4 GDF8 14260 17 7 6 64E 06 3 33E 06 50 18 2 2 4 GDF8 2 261 21 3 6 23E 06 2 10 06 33 61 2 0 4 GDF8 5192 17 0 6 17 06 2 76E 06 44 65 2 0 4 GDF8 1481262 20 8 5 75 06 3 00E 06 52 20 1 9 4 GDF8 62188 21 6 5 02E 06 3 7 06 79 06 1 6 4 GDF8 2263 15 3 4 77 06 1 14 06 23 93 1 5 4 GDF8 ite 264 18 9 4 28E 06 2 17 06 50 73 1 4 4 GDF8 12265 18 9 4 27E 06 1 15 06 26 86 1 4 4 GDF8 1814 2 0 1 56E 05 9 51 04 60 91 0 1 4 GDF8 1580205 1 7 1 36E 05 6 83E 04 50 42 0 0 4 GDF8 266 1 9 1 18 05 3 34E 04 28 43 0 0 4 GDF8 1580203 1 8 1 09E 05 3 49 04 32 09 0 0 100000 R19 1 7615 A B 94181 150 10881202 144 O it amp t180 ita 19 210185 4185 temas 1823 10 20 30 40 10 Ri DAA T WAA Wnt3a x 6 P gt SNI MU 0 0 781 1 563 3425 6 25 125 25 50 100 200 400 TIFEVAR E ng mL 5 1 73638 d AA 1 25 WAA Wnt3a A 20 Moo 15 St TR 10 N 5 0 y T T T T 1 5 9 0 781 1 563 3125 625 12 5 25 50 100 200 400 FIFEVARE ng mL 14 Data SI 17 amp 8 12 H e 08 06 2 lt
143. 8E 08 51 8 2 TT 8907 56 3 9 99EtO7 350 2 7 46 6 8 90E 07 31 2 2 8044 50 9 8 81E 07 2 EGF FGF PDGF A VEGF PDGF D A E L GDF8 1581028 7591 48 0 8 77 07 30 7 2 PDGF A VEGF PDGF D E L GDF8 4049 25 6 8 23E 07 28 8 2 PDGF A PDGF D GDF8 689133 7485 47 4 8 10E 07 28 3 2 EGF FGF PDGF A VEGF PDGF D 3 GDF8 8425 6571 41 6 7 60 07 26 6 2 EGF FGF PDGF A VEGF PDGF D GDF8 19182 7631 48 3 6 74E 07 23 6 2 FGF PDGF A VEGF PDGF D GDF8 8189 6777 42 9 5 93E 07 20 8 2 EGF FGF PDGF A VEGF PDGF D AY E L GDF8 184184 5475 34 6 5 44E 07 19 0 2 FGF PDGF A VEGF PDGF D E JL GDF8 8185 4093 25 9 4 92E 07 17 9 2 PDGF A PDGF D AZ E JL GDF8 1 542186 5274 33 4 4 68E 07 16 2 2 EGF FGF PDGF A VEGF PDGF D A E L GDF8 1 8 187 5342 33 8 4 2 7 14 1 2 EGF FGF PDGF A VEGF PDGF D lt GDF8 2 188 5533 35 0 3 98E 07 13 9 2 EGF VEGF PDGF D AY E L GDF8 1 8 189 5928 37 5 8 96E 07 13 9 2 PDGF A VEGF PDGF D E L GDFS 190 4822 30 5 3 90E 07 13 7 2 FGF PDGF A VEGF PDGF D A E JL 8191 4249 26 9 3 81E 07 13 3 2 EGF FGF PDGF A VEGF PDGF D A2 JL GDF8 15001092 5616 3
144. F D A E JL 8136 7964 59 5 9 47E 07 37 8 1 EGF FGF PDGF A VEGF PDGF D A2 JL GDF8 ite 4132 8066 60 2 9 29 07 37 1 1 EGF FGF PDGF A VEGF PDGF D GDF8 1681326 7415 55 4 8 30E 07 33 1 1 FGF PDGF A VEGF PDGF D GDF8 6994 52 2 7 76 07 31 1 EGF PDGF A VEGF PDGF D A1 JL GDF8 ite 31 6957 51 9 6 59E 07 26 1 EGF FGF PDGF A PDGF D AZ E JL GDF8 179 3573 26 7 2 07 9 7 1 TE 922 6 9 2 205807 1 0 1 2 68E 07 10 7 2 15494 98 0 1 25 07 44 2 FIFEYA Wnt3a 15807 100 0 2 86E 08 100 0 2 EGF FGF PDGF A VEGF PDGF D 3 GDF8 8818 21102 133 5 4 18E 08 146 3 2 EGF PDGF A PDGF D GDF8 158 15 15373 97 3 3 74E 08 130 8 2 FGF PDGF A VEGF PDGF D AZ E JL GDF8 814 9008 57 0 2 62E 08 91 6 2 EGF FGF PDGF A VEGF PDGF D Av E JL GDF8 124138 9650 61 0 2 46E 08 86 2 2 EGF FGF PDGF A VEGF PDGF D AY GDF8 1583235 10461 66 2 1 59 08 55 7 10 20 30 000 119 2011 526786 2011 10 20 ERRORS RI lo Sox1756H 7 ALE tamas vistas Btm 2 FGF PDGF A VEGF PDGF D A2E JL GDF8 816 9064 57 3 1 4
145. F PDGF D GDF8 3473 22 0 1 46 07 5 1 62110 PDGF A VEGF PDGF D 2 GDF8 3703 23 4 1 32 07 4 6 68111 EGF FGF PDGF A PDGF D 5 GDF8 2918 18 5 1 22E 07 4 3 112 EGF FGF PDGF A VEGF PDGF D AZ EJ GDF8 2975 18 8 1 04E 07 3 6 158 113 EGF FGF VEGF PDGF D A EJ GDF8 2910 18 4 9 18E 06 3 2 62114 FGF PDGF A VEGF PDGF D EJ GDF8 2734 17 3 6 13E 06 2 1 19115 EGF PDGF A VEGF PDGF D A EJ GDF8 2169 13 7 3 77 06 1 39 1858116 EGF FGF PDGF A VEGF PDGF D 22 GDF8 3107 19 7 3 52E 06 12 4639117 EGF FGF PDGF A VEGF PDGF D EJ GDF8 3343 21 1 5 35 05 0 2 08118 EGF FGF PDGF A VEGF PDGF D 4 GDF8 3034 19 2 2 37E 05 0 1 1589119 EGF FGF PDGF A VEGF PDGF D EJ GDF8 2263 14 3 1 66E 06 0 6 15891120 EGF FGF PDGF A VEGF PDGF D A1E JL GDF8 1771 11 2 5 57E 06 2 0 134121 PDGF A VEGF PDGF D 2 GDF8 1136 7 2 1 79 07 6 3 181122 PDGF A VEGF PDGF D Ai EJ GDF8 2021 12 8 2 07 7 3 10 20 114 2011 526786 2011 10 20 10 00 R5 7 5 An 3E ER 96 FIORIRE
146. GDF8 158 14 8464 53 5 8 33E 07 29 1 2 FGF PDGF A VEGF PDGF D A2E JL GDF8 6215 7234 45 8 7 95E 07 27 8 2 EGF FGF PDGF A VEGF PDGF D A2E JL GDF8 141100 6805 43 0 5 88E 07 20 6 2 101 5668 35 9 5 34E 07 18 7 2 FGF VEGF PDGF D GDF8 48102 6195 39 2 5 29 07 18 5 2 FGF PDGF A VEGF PDGF D Av GDF8 9103 7545 47 7 5 13 07 18 0 2 62104 4757 30 1 4 07 16 0 2 EGF FGF PDGF A VEGF PDGF D AY EJ GDF8 18105 6285 39 8 4 29 07 15 0 2 EGF FGF PDGF A VEGF PDGF D A1 JL GDF8 62106 5622 35 6 2 86E 07 10 0 2 FGF PDGF A VEGF PDGF D AZ E JL GDF8 184107 3951 25 0 1 72 7 6 0 2 EGF FGF PDGF A VEGF PDGF D A2 JL GDF8 194108 3226 20 4 1 58 07 2 EGF PDGF A VEGF PDGF D a GDF8 3473 22 0 1 46E 07 5 1 2 PDGF A VEGF PDGF D Av E L GDF8 110 3703 23 4 1 07 4 6 2 EGF FGF PDGF A VEGF PDGF D GDF8 8111 2918 18 5 1 22 07 4 3 2 FGF PDGF A VEGF PDGF D GDF8 62112 2975 18 8 1 04 07 3 6 2 FGF PDGF A VEGF PDGF D AZ E JL GDF8 158 113 2910 18 4 9 18E 06 3 2 E 10 20 30 000 118 2011 526786 2011 10 20
147. GF4 PDGF A VEGF AAT E Wnt3a amp 8 EGF FGF44 PDGF A A ZE JL 10071 96 30 2 95E4 06 1 85 158037 8 8 7966 76 17 1 19E 07 7 42 1585937 AAT Wnt3ae EGF FGF4 6932 66 28 4 62E 06 2 89 7 Wnt3a S EGF FGF4 PDGF AB 7473 71 46 2 61E 06 1 63 30 115 2011 526786 2011 10 20 ERRORE 5 75815 Fafa atta FIORIRE 519188 96 189337 AAT Wnt3as EGF FGF4 PDGF AB 456 E L 7914 75 67 1 91E 06 1 20 1585237 AAT Wnt3as EGF PDGFT A VEGF 12956 123 88 1 25E 07 7 82 Wnt3a dB FGF4 PDGF A VEGF 1 10E 07 897 AAT 38 Wnt3a amp EGF FGF4 PDGF A VEGF 1 05 1585537 AAT Wnt3a f EGF FGF4 PDGF A AZ JL 1 22 07 62934 2 78E 07 158 34 AAT 8 EGF FGF4 1 23E 08 41088184 AAT 8 8 EGF FGF4 PDGF AB 1 08 462 9134 AAT Wnt3a B EGF FGF4 PDGF AB t L E L 6 90E 06 1581534 AAT E Wnt3a S EGF PDGF A VEGF 4 04E 06 15434 AAT E FGF4 PDGF A VEGF 1 15 07 1084194 AAT EGF FGF4 PDGF A VEGF 5 01E 07 534 AAT Wnt3
148. OXA2 SOXI7 19 4 182 20 1 18 4 20 1 20 3 9 20 2 19 8 Wnt3a 15834181 158 180 19 4 8 202 40 GSK3i IXbio ES O CXCR4 FOXA2 SOX17 2 1 146 2011 526786 2011 10 20 40 2 ES 3 0 u 2 0 Ki g 1 0 90 D 8 E 5 2 amp m 5 2 SOX17 21 6 246 25 9 25 2 Wnt3a 212 23 6 257 250 desta 181 20 7 23 0 25 2 23 7 15 180 20 9 227 25 0 242 ds 19 196 218 242 226 202 18 5 22 4 236 223 desta 40 197 224 249 237 GSK3i IX bio 19 6 233 247 233 16833 56 19 7 23 0 25 3 24 4 25 x 20 8 15 5 3 40 05 lt 0 0 GAPDH CDX2 tal 18 0 23 8 25 5 Wnt3a 114 23 1 23 1 181 18 8 23 4 23 5 180 18 8 228 23 7 i815 19 183 220 250 158 202 18 7 238 23 0 40 18 4 23 0 23 2 SSK3i 184 222 240 156 56 18 2 23 8 23 3 2 5 2 ES 20 y 15 5 5 10 5 0 5 0 0 gt S y 45 4 lt 59 y Cox
149. RRORE RI RO 17 ZU HER tE E 163972 Zv hi 1873 26 53 2 93E 07 35 25 Zv hi 11874 2853 26 86 2 25E 07 27 01 Zv ki 41675 15 91 1 42E 07 17 05 12076 Zv F1 1585077 21 71 2 04E 07 24 55 20 01 134178 3298 31 06 2 58E 07 31 00 gt 3479 26 88 2 79E 07 33 47 FLA 33 93 3 22 7 38 62 Zv H Zv h 7 1 14484 2024 19 06 2 48E 07 29 80 7 1 85 2840 26 74 3 07 41 44 4 2986 1447 13 63 8 06 10 13 L H1 18487 5336 50 25 4 20E 07 50 38 27 PIFEMAT Wna 4033 35 50 2 14E 07 21 70 Zv k2 FIFCLA Wnt3a 4292 37 78 9 86E 07 100 00 ZL kF2 30 06 4 17E 07 42 28 Zv k2 188589 41 82 2 11E 07 21 40 7 2 409190 4542 39 98 8 7 30 7 Zv k2 2 2 L44992 37 06 2 95E 07 29 90 JZv Fk2 1t 89593 4157 36 59 2 29E 07 23 26 Zv F2 8794 4046 35 61 2 85E 07 28 91 L F2 8368 4 02E 07 L F2 t 96 3695 2 92E 07 Zv k2 15845397 3437 2 41E 07 Zv k2 L44998 4178 36 77 3 75 07 38 07 Zv k2 158599 3739 32 91 2 10ET 07 21 29 ZvL k2 20 02 1 27E 07 12 86 Zv k2 1E amp 93101 3496 30 77 2 98E 07 30 17 Zv k2 42 90 2 10 07 21 32 Zv k2 Zv k2 1 8 104 6115 53 82 4 93E 07 49 99 L FH2 15855105 6484 57 07 5
150. T amp f 00126 19961 223 24 5 75E 05 0 29 6235 8 amp 14238 159 24 1 85E 06 94 2 Wnt3a tA 6049 45 2 1 31 07 5 2 FIFUCVA Wnt3a 13392 100 0 2 50E 08 100 0 103120 EGF PDGF A VEGF PDGF D E L GDF8 9434 70 4 1 48E 08 59 1 EGF VEGF PDGF D 7988 59 6 1 13E 08 45 0 158 16 PDGF A PDGF D A JL GDF8 8303 62 0 9 20E 07 36 7 62913 FGF PDGF A PDGF D GDF8 7045 52 6 7 22E 07 28 8 PDGF A PDGF D AY E L GDF8 7799 58 2 6 82E 07 27 2 15492 FGF PDGF A VEGF PDGF D GDF8 5886 44 0 5 63E 07 22 5 FGF PDGF A VEGF PDGF D Ay E JL GDF8 5463 40 8 4 38E 07 17 5 FGF PDGF A VEGF PDGF D 2 GDF8 5100 38 1 4 18 07 16 7 1084195 PDGF A VEGF PDGF D 3 GDF8 4510 33 7 3 32E 07 13 3 83196 EGF FGF PDGF A PDGF D GDF8 4570 34 1 3 09E 07 12 3 FGF PDGF A VEGF PDGF D GDF8 4561 34 1 2 15 07 8 6 1598 FGF PDGF A VEGF PDGF D Av E L GDF8 3176 23 7 9 86E 06 3 9 108199 FGF VEGF PDGF D 5 GDF8 1209 9 0 1 56E 07 6 2 2
151. a EGF FGF4 PDGF A Ay E JL 5 56E 07 uu T2TULAT Wnt3ai 10224 108 14 7 36E 05 0 43 PIFEVA Wnt3a 9455 100 00 1 72 08 100 00 0 899 Wnt3a amp B 1 49 05 083139 AAT Wnt3agh EGF FGF4 5 11 06 AAT Wnt3a gA EGF FGF4 PDGF AB 1 62E 06 8499 AAT 8 f EGF FGF4 PDGF AB 4 5 E L 2 22E 06 1088199 AAT Wnt3a EGF PDGF A VEGF 4 84E 06 58939 AAT 4 8856 93 67 7 01 05 0 41 083139 AAT Wnt3agh EGF FGF4 PDGF A VEGF 5 30E 06 Wnt3a 8 EGF FGF44 PDGF A A ZE JL 7 32E4 05 15422 Wnt3agh EGF 1 75E 06 8422 AAT 8 f EGF FGF4 1 04E 07 158 22 AAT 3 8 EGF FGF4 PDGF AB 8 60E 06 14422 AAFT WntSadB EGF FGF4 PDGF AB A ZE JV 14676 155 23 5 612 06 3 27 8022 AAT Wnt3a EGF PDGF A VEGF 20372 215 48 4 99E 06 2 91 462 22 8 Wnt3a amp FGF4 PDGF A VEGF 12277 129 85 4 6 2 86 8422 AAT EGF FGF4 PDGF A VEGF 12522 132 44 7 88 6 4 59 28422 AAT amp Wnt3a SB EGF FGF4 PDGF A LA EJ 11610 122 80 1 33E 07 7 77 Aa DUH E 8 1 73635 TE TEXE 96 773 3 3 477 6 64 7 AE 04 0 09 FIFEVA Wint3a 7185 100 00 8 07 100 00 AARS Wnt3ad f ZU 4611 64 18 1 4 07 17 21 AAT Wnt3a
152. a 6 6049 45 2 1 31E 07 5 2 PA4FEVA Wnt3a 13392 100 0 2 5 100 0 FGF PDGF A VEGF PDGF D 2 GDF8 1 18 13037 97 3 1 63E 08 65 2 EGF FGF PDGF A VEGF PDGF D A2 JL GDF8 ite 9914 9344 69 8 1 23E 08 49 0 FGF PDGF A PDGF D E L GDF8 158 15 8448 63 1 8 64 7 34 5 FGF PDGF A PDGF D Av E L GDF8 583116 5498 41 1 6 56E 07 26 2 FGF PDGF A VEGF PDGF D Li EJ GDF8 564 5063 37 8 5 88E 07 23 5 EGF FGF PDGF A VEGF PDGF D GDF8 583165 4788 35 8 4 57 7 18 2 FGF PDGF A VEGF PDGF D 99 E L GDF8 494166 8129 60 7 3 53E 07 14 1 EGF FGF PDGF A VEGF PDGF D 5 GDF8 449167 6791 50 7 8 18 07 12 7 PDGF A VEGF PDGF D 5 GDF8 124468 3456 25 8 2 30E 07 9 2 ameo 3995 29 8 1 69E 07 6 8 4 FGF PDGF A VEGF PDGF D 55 GDF8 474 3 5 1 80E 07 7 2 Wnt3a ZL 15494 98 0 1 25 07 4 4 2 2 POIFEVA Wnt3a TEL 15807 100 0 2 86E 08 100 0 2 EGF FGF PDGF A VEGF PDGF D 3 GDF8 1688119 8425 53 8 1 19E 08 41 6 2 EGF FGF PDGF A PDGF D 5 GDF8 9123 57 7 1 13E 08 9 7 2 PDGF A PDGF D 4 tE L GDF8 8471 6048 38 3 5 5
153. er special reason as specified document referring to an oral disclosure use exhibition or other means P document published prior to the international filing date but later Ihan the priority date Date of the actual completion of the international search 30 October 2009 Name and mailing address of the ISA European Patent Office 5818 Patentlaan 2 NL 2280 HV Rijswijk Tel 31 70 340 2040 Fax 431 70 340 3016 Form 5 210 second sheet 2005 Invention X document of particular relevance the claimed invention cannot be considered novel or cannot be considered to Involve ari inventive step when the document is taken alone Y document of particular relevance the claimed Invention cannot be considered to involve an inventive step when the documen is combined with one or more other such docu ments such cornbinalion being obvious to a person skilled in the art document member of the same patent family Date of mailing of ihe intemalional search report 11 11 2009 Authorized officer Brero Alessandro 152 2011 526786 2011 10 20 International application No _____ PCT US2009 049183 Citation of document with indication where appropriate of the relevant passages Relevant ta claim No INTERNATIONAL SEARCH REPORT WO 2008 048647 A CYTHERA INC US BAETGE EMMANUEL EDWARD US D AMOUR KEVIN US
154. msa JL F6 135 10068 213 2 15E 06 0 68 20 06 18436 5170 109 39 1 22E 06 0 39 Zv k6 1584237 65 55 1 91E 06 0 61 20 30 40 109 2011 526786 2011 10 20 ERRORE Ei 17 319 3085 2 88 L F6 4H38 1537 32 52 4 48E 04 0 01 gt 9039 3650 77 23 2 01E 06 0 64 L F6 14840 5817 123 07 4 91 05 0 16 7 66 1 4 92 23 1 07 05 0 03 JL F6 4035 85 38 2 06 0 66 7 6 865 3279 69 37 5 6 05 0 18 J L F6 4 2 67 2698 57 08 1 95E 06 0 62 2 07 FIFUVAT Wnt9a amp 9545 321 22 4 87E 06 0 99 7270 07 72 ELAZWnt3a 20064 675 20 4 92E 08 100 00 Zv k7 121168 366 62 5 15E 06 1 05 Zv k7 185569 327 58 8 97E 06 0 81 L F7 1653170 563 21 6 51E 06 1 32 ZU k7 605 71 7 06 1 50 Zv k7 245 96 6 7 6 1 32 7 7 8173 3 03E 06 L F7 974 2 67E 06 70 07 975 7 75E 06 Zv k7 76 485 89 4 19 06 0 85 2 414 95 6 03 06 1 22 Zv k7 185579 8535 287 22 6 92bET O6 1 41 ZL k8 295 49 2 1 06 0 52 Zv k8 799 17 4 08 100 00 JZv F8 125580 13939 599 68 4 23E 06 0 95 7 8 L44981 10466 450 29 4 91E 06 1 10 ZL F8 1482 10323 444 14 4 90E 06 1 10 7 8 88
155. o 10 October 2007 2007 10 pages 2419 2430 XP002547893 ISSN 1076 3279 the whole document Form POTASA 210 continuation of second shoot April 2005 153 2011 526786 2011 10 20 International application No PCT US2009 049183 Citation of document with indication where appropriate of the relevant passages INTERNATIONAL SEARCH REPORT Relevant to No SHIRAKI NOBUAKI ET AL Guided differentiation of embryonic stem cells into Pdxl expressing regional specific definitive endoderm 0 STEM CELLS DAYTON OHIO APR 2008 vol 26 no 4 April 2008 2008 04 pages 874 885 XP002547894 ISSN 1549 4918 the whole document KUBO A ET AL Development of definitive endoderm from embryonic stem cells culture DEVELOPMENT COMPANY OF BIOLOGISTS CAMBRIDGE 68 vol 131 no 7 1 April 2004 2004 04 01 pages 1651 1662 002985523 ISSN 0950 1991 th whole document Form PCT ISA 210 continuation second sheet April 2005 154 2011 526786 2011 10 20 INTERNATIONAL SEARCH REPORT E international application No PCT US2009 049183 Git d in search report date member s date WO 2008048647 24 04 2008 information on patent family members Forn PCTASA 210 pateni family annex April 2005 155 2011 526786 2011 10 20 1000000000
156. ymbols 6128 Documentation searched other than minimum documentation the extent that such documents are included in the fields searched Electronic data base consulted during the Intemational search name of data base and where practical search terms used EPO Internal BIOSIS EMBASE WPI Data DOCUMENTS CONSIDERED TO BE RELEVANT Citation of document with indication where appropriate of the relevant passages Relevant to claim No MCLEAN AMANDA B ET AL Activin a efficiently specifies definitive endoderm from human embryonic stem cells only when phosphatidylinositol 3 kinase signaling is suppressed STEM CELLS ALPHAMED PRESS DAYTON 0H US vol 25 no 1 1 January 2007 2007 01 01 pages 29 38 XP002482637 ISSN 1066 5099 abstract 3 page 32 page 33 figure 3 x Further documents are listed in the continuation of Box C x See patent family annex Special categories of citad documents later document published after the international date or priority date and not in conflict wilh the application but A document defining the general state of the art which la noi cited to understand the principle or theory undertying the considerad lo be of particular relevance aarller document but published on or after he international filing date L document which may throw doubts on priority claims or which is cited lo establish the publicatlon date of another citation or olh
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