Figure 2.

Sox2-low iPSCs do not self-renew in serum plus LIF

(A) Phase and Rex1-GFP images of Sox2^−/− rMKOS (−/− Sox2-low) iPSCs and ESCs at passage 0 after medium switch from 2i plus LIF into serum plus LIF.

(B and C) qRT-PCR analysis of pluripotency markers (B) and trophoblast markers (C) 3 days after switching into serum plus LIF medium.

(D) Western blot of Sox2-low iPSCs in 2i plus LIF with and without rescue Sox2 transgene (+Sox2). Rex1-GFP WT iPSCs were used as control for Sox2 protein levels.

(E) Phase and GFP images of −/− Sox2-low + Sox2 rescue iPSCs after 5 passages in serum plus LIF.

(F) qRT-PCR analysis, after 3 days in serum plus LIF, of pluripotency factors and Pl-1 in −/− Sox2-low iPSCs with and without a rescue Sox2 transgene. ESCs were provided as control in the qRT-PCR analysis of pluripotency factors.

Scale bars = 200μm. Error bars indicate standard deviation of replicate qPCR reactions (n = 3).