Figure 3.

Sox2-low and null nPSCs differentiate into extraembryonic and all embryonic lineages

(A) qRT-PCR analysis of embryoid body assay using Sox2^−/− (−/−) Sox2-low iPSCs, control iPSCs (WT) and ESCs showing RNA expression of markers of pluripotency (Nanog, Oct4), ectoderm (FGF5), mesoderm (T Brachyury and Zeb2), endoderm (FoxA1 and Gatat4) and trophoblast (Pl-1 and Elf5).

(B) qRT-PCR analysis of embryoid body assay for retroviral Sox2 expression in −/− Sox2-low iPSCs.

(C) qRT-PCR analysis of embryoid body assays of −/− Sox2-low and rescue (+Sox2) iPSCs, for expression of pluripotency (Nanog), late epiblast (FGF5) and trophectoderm (PL-1) markers.

(D) Western blot showing Sox2 protein post tamoxifen (4OHT) treatment in Sox2^FLIP/FLIP ESCs with or without a constitutive CreERT2 transgene. Carrier only (ethanol, ETOH) and Sox2^+/+ ESCs were used as additional controls.

(E) Experimental design of embryoid body assay and of Sox2 deletion in Sox2^FLIP/FLIP ESCs.

(F) qRT-PCR analysis of embryoid body assay using Sox2^FLIP/FLIP ESCs showing expression of pluripotency markers (Nanog), late epiblast (FGF5), mesoderm (T Brachyury), endoderm (Gata4), trophoblast (Pl-1 and Elf5). Pl1 (∗) chart omits Sox2^FLIP/FLIP CreERT2 ESC sample treated with 4OHT.

Error bars indicate standard deviation of replicate qPCR reactions (n = 3).