Fig. 6: The PHD2/HIF-1α axis regulated mesendoderm through the Wnt/β-catenin pathway.

a The relative TOPflash activities in the WT, PHD2-KO/Ctrl, and PHD2-KO/PHD2-OE groups (n = 4). b The relative TOPflash activities in the Ctrl, HIF-1α, HIF-1α + PHD2 groups. (n = 4). c Western blots showing the protein expressions of key components of the Wnt/β-catenin pathway in the WT, PHD2-KO/Ctrl, and PHD2-KO/PHD2-OE mESCs, respectively. (n = 4). d The relative levels of β-catenin and p-GSK3β(S9) in (c) were quantified. e Western blots showing the protein expressions of key components of the Wnt/β-catenin pathway in the Ctrl, HIF-1α, HIF-1α + PHD2 mESCs, respectively. (n = 4). f The relative levels of β-catenin and p-GSK3β(S9) in (e) were quantified. g The peaks of the ChIP-seq against HIF-1α, H3K4me3, H3K27ac, H3K9me2, and H3K27me3 in mESCs at the locus of Ctnnb1. h The immunostaining against β-catenin in PHD2+/+ (n = 3), PHD2+/− (n = 5), and PHD2−/− embryos (n = 3) at E7.5 stage. i The fluorescence signal spectrum and j positive percentages of β-catenin in (h) were measured. Scale bars, 100 μm; WT, wild type; KO, PHD2-KO; OE, PHD2-OE; Ctrl, control; HIF-1α, HIF-1α overexpression; HIF-1α + PHD2, HIF-1α/PHD2 dual overexpressions; +/+ PHD2+/+, +/− PHD2+/−, −/− PHD2+/−, A anterior, P posterior; *P < 0.05; **P < 0.01; ****P < 0.0001.