hESdFs promote a sequential endodermal-to-hepatic differentiation in human and monkey ESCs. A, QRT-PCR analysis of IVDS 1, 2, and 3 for pluripotency-related markers (Oct4 and Nanog), mesodermal markers (Brachyury MIXL1), endodermal markers (SOX17, GATA4, and CXCR4), ectodermal markers (SOX1 and PAX 6), and hepatocyte markers (AFP, CK8, CK18, albumin, and CYP7A1). B, the expression of Oct4 (pluripotency-related), early ectodermal marker SOX1, early mesodermal Brachyury, and early endodermal SOX17 in differentiating human and monkey ESC colonies of IVDS 1 and 2 were analyzed by ICC. C, co-expression of endodermal marker SOX 17 with hepatocyte marker CK18 or AFP is as indicated. IVDS2 foci derived from monkey ESC were stained for SOX17 (red) and CK18 (green) or AFP (green). Scale bar, 25 μm.