Fig. 6

MYH1G-AS interacts with FGF18 to destroy FGF18 protein stabilization. A Venn diagram showing the specific binding proteins of the MYH1G-AS sense strand or antisense strand. B, C The interaction of MYH1G-AS with FGF18 protein was determined by biotin-labeled RNA pulldown (B) and RIP (C) assays. D The binding of full-length and truncated MYH1G-AS with FGF18 protein was determined by RNA pulldown assay. E IF staining of FGF18 in CPM. F, G The protein expression level of FGF18 after MYH1G-AS overexpression or knockdown in vitro (F) and in vivo (G). H Left: FGF18 protein expression in myoblasts after dimethyl sulfoxide (DMSO) or cycloheximide (CHX; 25 µg/mL) treatment for 12 h. Right: FGF18 protein expression in the MYH1G-AS-knockdown myoblast was analyzed after incubation with CHX. I Left: FGF18 protein expression in myoblasts after DMSO or MG-132 (5 µmol/L) treatment for 12 h. Right: FGF18 protein expression in MYH1G-AS overexpressed myoblast was analyzed after incubated with MG-132. J−O Relative mRNA expressions of several cell cycle-inhibiting genes (J), relative mRNA expressions of myoblast differentiation marker genes (K), relative mtDNA content (L), mitochondrial membrane potential (M), [ROS]i (N), and relative mRNA expressions of FBXO25 (O) induced by the listed nucleic acids in CPMs. In panel F−I, the numbers shown below the bands were folds of band intensities relative to the control. Band intensities were quantified by ImageJ and normalized to β-tubulin. Results are presented as mean ± SEM. In panels J–O, the statistical significance of differences between means was assessed using an independent sample t-test. (*P < 0.05; **P < 0.01)