Fig. 7: miR-429 is an upstream regulator of PHD2 in mesendoderm differentiation.

a The RNA levels of miR-429 along the mesendoderm differentiation of mESCs under normoxia and hypoxia, respectively. (n = 4). b The RNA levels of miR-429 and Phd2 along the mesendoderm differentiation. (n = 4). c Western blots showing the PHD2 expression change with miR-429 overexpression. (n = 3). d The luciferase assays were performed to prove miR-429 targeting Phd2 mRNA. (n = 4). e The immunostaining against T and f the T-positive cell ratios after the mesendoderm differentiation of the ctrl and miR-429-OE mESCs. Blue region represents gating negative cells and red region represents treated cells. (n = 4). g The schematic diagram showing the construction of Dox-inducible miR-429/PHD2 dual overexpression (429-P-OE) mESCs. h The overexpression of PHD2 and i miR-429 in 429-P-OE were verified. j The immunostaining against T and k the T-positive cell ratios after the mesendoderm differentiation of the Ctrl and miR-429-OE mESCs (n = 4). l In the presence of abundant oxygen, a working model of miR-429, PHD2, HIF-1α, and the Wnt/β-catenin pathway in regulating mesendoderm specification. Scale bars, 100 μm; ctrl/Ctrl no inducible control; miR-429-OE miR-429 overexpression, 429-P-OE miR-429/PHD2 dual overexpressions; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns not significant.