Fig. 5.

Hypoxia treatment after the mesendoderm promoted the differentiation of the mesendoderm-derived lineages. A Schematic diagram of the 8-day AB2.2 mESC differentiation that consisted of a 4-day differentiation under normoxia and a following 4-day differentiation under either normoxia or hypoxia. B Wnt3, Wnt8a, C Sp5, and Cdx1 were all significantly repressed by hypoxia. D Cardiac markers (Tbx5, Mef2C, Nkx2-5, and α-MHC) were significantly upregulated by hypoxia. E Pecam1 and Acta2 were significantly upregulated by hypoxia. F By immunostaining on differentiation day 8, the expression of α-Actinin (red) was shown to be enhanced. Nuclei were stained with DPAI (blue). G By immunostaining on differentiation day 8, the expression of α-SMA (red) was shown to be enhanced. Nuclei were stained with DPAI (blue). H The ratios of cTnT+ and α-SMA+ cells on differentiation day 8 in AB2.2 mESCs were detected by flow cytometry. *, Significant (P<0.05)