Figure 4.

The reverse effects of METTL3 over-expression on the anti-hypertrophy effect of MA in vivo. (A) The effect of METTL3 over-expression on the MA impaired total RNA m⁶A methylation content in TAC induced hypertrophic LV tissues detected with the EpiQuik m⁶A RNA Methylation assay kit and (B) confirmed by Dot Blot. (C) The ratio of HW/BW and HW/ TL shows the effect of METTL3 over-expression on the MA inhibited morphology of cardiac morphology. (D) Analysis of the effect of METTL3 over-expression on the MA preserved cardiomyocyte area in left ventricle through Histology H&E and (E) WGA staining of cardiac cross-sections. (F) Echocardiography detection of the parameters of LVEF, LVFS, LVEDV, LVESV, LVEDD, and LVESD to evaluate the effect of METTL3 over-expression on the MA preserved cardiac function of TAC mice. (G) Real-time PCR analysis of the mRNA levels of the hypertrophy markers (ANP, BNP, and β-MHC). (H) Western blot detection of the protein levels of the hypertrophy markers (ANP, BNP, and β-MHC). (I) Detection of the effect of MA on the TAC-induced myocardial fibrosis via Masson staining. (J) Real-time PCR analysis of the mRNA levels of the myocardial fibrosis (Fibronectin, Collagen I, and α-SMA). (K) Western blot analysis of the protein levels of the myocardial fibrosis (Fibronectin, Collagen I and α-SMA); *P<0.05, ** P <0.01 compared to the indicated group.