Fig. 5: HIF-1α mediated the effects of PHD2 on mesendoderm specification.

a The immunostaining against HIF-1α and PHD2 in PHD2+/+ (n = 4), PHD2+/− (n = 6), and PHD2−/− embryos (n = 3) at E7.5 stage. b The fluorescence signal spectrum and (c) positive percentages of HIF-1α in (a) were measured. d Western blots showing the protein expressions of HIF-1α in the WT and PHD2-KO mESCs that were treated with either DMSO or IOX4 (n = 4). e The relative levels of HIF-1α in (d) were quantified. f The mRNA levels of Pgk1 and Ldha after the mesendoderm differentiation of the WT and PHD2-KO mESCs that were treated with either DMSO or IOX4. (n = 4). g Western blots showing the protein expressions of HIF-1α and PHD2 in the mESCs of the Ctrl, HIF-1α, and HIF-1α + PHD2 groups, respectively. (n = 4). h The immunostaining against T and i the mRNA levels of mesendoderm markers after the mesendoderm differentiation of mESCs of the Ctrl, HIF-1α, and HIF-1α + PHD2 groups, respectively (n = 4). Scale bars, 100 μm; +/+ PHD2+/+, +/− PHD2+/−, −/− PHD2+/−, A anterior, P posterior, WT wild type, KO PHD2-KO, DMSO DMSO treatment, IOX4 IOX4 treatment, Ctrl control, HIF-1α HIF-1α overexpression; HIF-1α + PHD2, HIF-1α/PHD2 dual overexpressions; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns not significant.