Pcgf1 regulated neural induction through histone methylation. (A) qPCR analysis of the expression levels of BMP signaling pathway (Smad1, Smad4, and Smad5) and Wnt signaling pathway (wnt3a, wnt8a, and β-catenin). Compared with the controls, the mRNA levels of Smad1, Smad4, and Smad5 in Pcgf1 MO-injected embryos decreased, but the levels of wnt3a, wnt8a, and β-catenin did not change observably. Data represent the mean of at least three independent experiments ± SD. *P < 0.05 vs. control. (B) Compared with the embryos injected with Pcgf1 MO, the shrinking of the telencephalon has not been rescued by the addition of Smad4 mRNA. (C) Western blot showed that the expression of H3K4me3 and H3K27me3 decreased after Pcgf1 knocked down. (D) The WT and Pcgf1 MO-injected group were immunoprecipitated with anti-H3K27me3, anti-H3K4me3, and IgG. The isolated DNA was analyzed by gene-specific ChIP primers. The levels of H3K27me3 at the promoters of Ngn1 and Otx2, and the levels of H3K4me3 at the promoters of Pou5f3 and Nanog, were significantly decreased after injection with the Pcgf1 MO. Data represent the mean of at least three independent experiments ± SD. *P < 0.05, **P < 0.01.
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