FIGURE 3.

Apol9a knockdown inhibits C2C12 myogenesis. C2C12 cells were induced to differentiate after si-NC or si-Apol9a transfection for 4 days. (A) qPCR and western blot validation of the efficiency of Apol9a knockdown in C2C12 cells. (B) Protein expression levels of MyHC, MyoD, Myf5, and MyoG were estimated through western blot analysis after si-NC and si-Apol9a transfection. (C) Gray scale analysis of western blot results of (B) determined using ImageJ software. (D) qPCR was performed to assess the mRNA expression levels of several myogenic differentiation genes (Myf5, MyoD, MyoG, and MyHC). (E) Myotube formation can be observed in fluorescence images of DAPI (blue) and MyHC antibody-stained (green) myotubes of C2C12 cells transfected si-NC or si-Apol9a. Scale bar, 100 μm. The fusion index (%) was quantified and is shown on the right. The data are expressed as mean ± SEM of at least three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 vs. si-NC control.