Fig. 3

METTL3 was involved in regulating cellular decidualization and embryo implantation. a, b Overexpression and knockdown (ovM3, siM3) efficacy of METTL3 at the mRNA level in human endometrial stromal cells (ThESCs). c, d Overexpression and knockdown efficacy of METTL3 at the protein level in ThESCs. e, f Dot blot assays were used to detect m6A changes upon METTL3 overexpression or knockdown in ThESCs. Methylene blue staining was performed as a control. g-j Protein levels of PRL and IGFBP1 upon METTL3 overexpression or knockdown in ThESCs. k, l mRNA and protein levels of FOXO1 upon METTL3 overexpression in ThESCs. m, n mRNA and protein levels of FOXO1 upon METTL3 knockdown in ThESCs. o Cell percentage of HTR-8/SVneo trophoblast cell line spheroids attached to ThESCs following METTL3 overexpression or knockdown were evaluated (scale bar = 200 μm). p Protein level of FOXO1 reduced by METTL3 overexpression was rescued by FOXO1 overexpression. q Protein level of FOXO1 increased by METTL3 knockdown was decreased by a FOXO1 inhibitor. r Cell percentage of HTR-8/SVneo spheroids attached to ThESCs transfected with the negative control and METTL3-overexpressing vector were evaluated in the absence or presence of the FOXO1-overexpressing vector. The cell percentage of HTR-8/SVneo spheroids attached to ThESCs transfected with the negative control and siRNA of METTL3 were evaluated in the absence or presence of a FOXO1 inhibitor (scale bar = 200 μm). ThESCs were treated with METTL3 intervention for 24 h and then subjected to decidual treatment with MPA + 8-br-cAMP for 4 d. 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 < 0.05, **P < 0.01, ***P < 0.001