Fig. 3.

Gata6, Gata4, Sox17 and Pdgfra are required for cXEN establishment. (A) mESCs mutant for Sox17, Gata4, Gata6, Pdgfra or Gata4 and Gata6 double-mutant cells were subjected to the cXEN protocol along with wild-type or Nanog overexpressing (OE) mESCs. Representative phase-contrast images were taken at defined time points during the course of derivation [day 0 in pluripotency conditions, day 3 prior to the first passage and the endpoint: after prolonged culture (day 30)]. Scale bars: 100 μm. (B) Quantification of Sox7 and Nanog expression following immunofluorescence analysis at 5 days and 30 days after cXEN derivation. Data are mean±s.e.m. of five separate counts. ^*P<0.05; ^**P<0.01; ^***P<0.001.