Fig. 1.

Hypoxia dramatically suppressed mesendoderm differentiation of mESCs. A Schematic diagram of mESC differentiation under normoxia or hypoxia. B Hypoxia significantly repressed the mRNA expression of mesendoderm markers (T, Eomes, Mesp1, and Gsc) in differentiating AB2.2 mESCs. C Hypoxia repressed T protein expression in differentiating AB2.2 mESCs. T (red) was stained on differentiation day 4. Nuclei were stained with DPAI (blue). D Hypoxia significantly upregulated the mRNA expression of ectoderm markers (Pax6 and Nestin) in differentiating AB2.2 mESCs. E Hypoxia promoted Sox1 protein expression in differentiating AB2.2 mESCs. Sox1 (green) was stained on differentiation day 4. Nuclei were stained with DPAI (blue). F The ratios of T+ and Sox1+ cells on differentiation day 4 ofAB2.2 mESCs were detected by flow cytometry. G Hypoxia significantly repressed the mRNA expression of cardiac markers (Tbx5, Mef2C, Nkx2.5, and α-MHC) in differentiating AB2.2 mESCs. H Hypoxia inhibited T protein expression in R1 mESCs. T (red) was stained on differentiation day 4. Nuclei were stained with DPAI (blue). I The ratios of T+ and Sox1+ cells on differentiation day 4 of R1 mESCs were detected by flow cytometry. *, significant (P<0.05)