Fig. 2

METTL3 mediate m6A methylation inhibits the progression of EMs by promoting cellular senescence in vitro and in vivo. A The measurement of the knockdown efficiency of si-METTL3 transfected ESCs in mRNA level by RT-qPCR. B The global m6A levels in si-METTL3 transfected ESCs detected by m6A dot blot assay with anti-m6A antibody. The increasing ability of viability, proliferation, invasion, and migration of ESCs with si-METTL3 transfection was detected by CCK8 C, EdU staining D), wound healing E and Transwell F assay. G The mRNA level of METTL3 in oe-METTL3 transfected ESCs by RT-qPCR. H The mRNA N6-methyladenosine level in oe-METTL3 transfected ESCs by m6A dot blot. I–L CCK8, EdU, wound healing, and Transwell assay showed the decreasing ability of viability, proliferation, invasion, and migration of ESCs with oe-METTL3 transfection in reverse. M Level of cellular senescence in ESCs transfect with si-NC, si-METTL3, oe-vector and oe-METTL3 by SA-β -Gal staining. N The protein level of METTL3 and senescence biomarkers in ESCs transfect with si-NC, si-METTL3, oe-vector, and oe-METTL3 by western blotting, using GAPDH as an internal control. O Mass morphology of AAV-shNC and AAV-shMETTL3 mice. P The xenografts were isolated and measured by caliper in AAV-shNC and AAV-shMETTL3 mice. Xenografts’ weight Q and volume R were measured and analyzed. S Representative staining images for cellular senescence biomarkers (Lamin b1, p53, and p21) in AAV-shNC(up) and AAV-shMETTL3 group(down) by IHC. (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