Fig. 7.

Hif-1α overexpression promoted the mesendoderm differentiation but repressed the ectoderm differentiation. A Hif-1α overexpression upregulated T protein expression under normoxia. T (red) was stained on differentiation day 4. Nuclei were stained with DPAI (blue). B Hif-1α overexpression induced by Dox supplementation repressed the expression of ectoderm markers (Pax6 and Nestin), although the difference in Nestin levels was not significant. C TOPFlash assays showed that Hif-1α knockdown significantly repressed the Wnt/β-Catenin pathway activation. D TOPFlash assays showed that Hif-1α overexpression significantly promoted the Wnt/β-Catenin pathway activation. E, F The expressions of Hif-1α in Hif-1α-iOE AB2.2 mESCs treated with 1, 10, 100, and 1000 ng/mL Dox, respectively. Normoxic and hypoxic cultures without dox treatment were used as controls. G Schematic diagram of Hif-1α-iOE AB2.2 mESC differentiation treated with 1, 10, 100, and 1000 ng/mL Dox, respectively. Normoxic and hypoxic cultures without dox treatment were used as controls. H The expression changes of mesendoderm markers (T, Eomes, Mesp1, and Gsc) on day 4 of the differentiation described in G. I The ratios of T+ and Sox1+ cells on day 4 of the differentiation assays in G were detected by flow cytometry. scramble, AB2.2/scramble cells; shHif-1α, AB2.2/shHif-1α cells; DOX, doxycycline; *, significant (P<0.05)