Fig. 6.

Hif-1α knockdown repressed the mesendoderm differentiation and promoted the ectoderm differentiation. A, B The expression patterns of Hif-1α in differentiating AB2.2 cells under normoxia and hypoxia. C Schematic diagram of AB2.2 mESC differentiation treated with normoxia and hypoxia on differentiation days 0–2 and 2–4, respectively. D The expression of mesendoderm markers (T, Eomes, Mesp1, and Gsc) was repressed to the same extent by hypoxia on differentiation days 0–2 and 2–4. E, F Hif-1α knockdown was verified by western blotting analysis under hypoxia. G Hif-1α knockdown inhibited T protein expression under hypoxia. T (red) was stained on differentiation day 4. Nuclei were stained with DPAI (blue). H Hif-1α knockdown promoted the expression of ectoderm markers (Pax6 and Nestin). I Schematic diagram of the parallel comparison of Hif-1α knockdown effects on AB2.2 mESC differentiation under either normoxic or hypoxic conditions. J Hif-1α knockdown inhibited the expression of mesendoderm markers (T, Eomes, Mesp1, and Gsc) on differentiation day 4 under both normoxia and hypoxia, with the effect more obvious under normoxia. The ratios of K T+ and L Sox1+ cells on day 4 of the differentiation assays in I were detected by flow cytometry. scramble, AB2.2/scramble cells; shHif-1α, AB2.2/shHif-1α cells; *, significant (P<0.05); ns, not significant