Fig. 3

SIRT1 as a downstream target and negatively regulated by METTL3-mediated m6A modification in vitro and in vivo. A Volcano plots for differentially expressed mRNAs between matched oe-vector and oe-METTL3 transfected ESCs by RNA-seq. (|Fold Change|≥ 1, p < 0.001, Red point: up-regulated mRNAs, blue point: down-regulated mRNAs, the grey point indicated not differential expressed). B Volcano plots for differential methylated m6A modification transcripts between matched oe-vector and oe-METTL3 transfected ESCs by m6A epitranscriptomic microarray.(|Fold Change|≥ 1). Hyper-Up, up-regulated m6A modified mRNAs of |Fold Change|≥ 2; Hyper-Down, down-regulated m6A modified mRNAs of |Fold Change|≥ 2; Hypo-Up, up-regulated m6A modified mRNAs of |Fold Change|< 2; Hypo-Down, down-regulated m6A modified mRNAs of |Fold Change|< 2. C Venn showed SIRT1 at the intersection of RNA-seq and m6A epitranscriptomic microarray. D Top results of KEGG enrichment analysis. KEGG, Kyoto Encyclopedia of Genes and Genomes. E The m6A motif of ESCs identifified by m6A epitranscriptomic microarray. F m6A peak distribution of mRNA in m6A epitranscriptomic microarray. G The decreasing SIRT1 expression level of oe-METTL3 transfected ESCs in RNA-seq. H The increasing m6A mythylation level of oe-METTL3 transfected ESCs in m6A epitranscriptomic microarray. I The mRNA level of SIRT1 in EMs and paired tissues by RT-qPCR. J Representative IF staining of METTL3 and SIRT1 expression in Ectopic, Eutopic, and normal endometrium tissues. (Red fluorescence: METTL3; Green fluorescence: SIRT1; Blue fluorescence: DAPI; scale bar: 100 μm). K mRNA level of SIRT1 in METTL3 knockdown(up) and overexpression(down) by RT-qPCR. L Reduction enrichment level of SIRT1 in ESCs after METTL3 silencing by RIP-qPCR. M Decreasing m6A modification level of SIRT1 transcripts during METTL3 knockdown by Merip-qPCR assay. N Workflow of Endometriosis CKO mice model. O Progression of ectopic lesions (Green rectangular range) on Days 3, 5, and 7 in control and CKO-METTL3 mice. The mRNA expression of METTL3 P and volume on Days 3, 5, and 7 Q of isolated ectopic lesions were measured and analyzed. R Representative IHC images for SIRT1 in Control(up) and CKO-METTL3 group(down). (scale bars = 100 μm and 50 μm). All above result were shown in means ± SD, ns, p ≥ 0.05; *p < 0.05; **p < 0.01; ***p < 0.001