Fig. 5
METTL3-mediated m6A regulated the degradation of FOXO1 mRNA in a YTHDF2-dependent manner. a The curve and statistical analysis of the FOXO1 mRNA decay slope in the negative or METTL3-overexpressing ThESCs after transcriptional inhibition. b RNA immunoprecipitation-PCR (RIP-PCR) assays showing an enrichment of FOXO1 bound to METTL3 in ThESCs. c Methylated RNA immunoprecipitation-PCR (MeRIP-PCR) assays showing an enrichment of FOXO1 with m6A in METTL3-overexpressing ThESCs. d Protein levels of METTL3 in the wild METTL3-overexpressing (ovM3-WT) and mutated METTL3-overexpressing (ovM3-MUT) ThESCs were analyzed by western blotting. e Methylation of FOXO1 mRNA in the ovM3-WT and ovM3-MUT ThESCs were analyzed by MeRIP-PCR. f mRNA levels of FOXO1 in the ovM3-WT and ovM3-MUT ThESCs were analyzed by qRT-PCR. g Protein levels of FOXO1 in the ovM3-WT and ovM3-MUT ThESCs were analyzed by western blotting. h Curve and statistical analysis of the FOXO1 mRNA decay slope in the ovM3-WT and ovM3-MUT ThESCs after transcriptional inhibition. i Enrichment of FOXO1 mRNA bound to YTHDF2 in the ovM3-WT and ovM3-MUT ThESCs were analyzed by RIP-PCR. j Model of a pattern of METTL3-mediated m6A in regulating the decidualization of endometrial stromal cells: METTL3 increases the m6A level of FOXO1 mRNA, thus promoting the binding of YTHDF2 and enhancing the degradation of FOXO1 mRNA, contributing to the defective decidualization of endometrial stromal cells in endometriosis. All experiments were repeated in triplicate or quadruplicate. The blots in this figure are cropped (please refer to supplementary files for details). Data with error bars are presented to indicate the mean ± SEM values. ***P < 0.001, ****P < 0.0001