Fig. 4: KAP1 complex regulates C2C12 myoblasts differentiation via miR-133a repression.

a KAP1 regulates the differentiation of C2C12 cells. The expression of Kap1 in C2C12 cells was manipulated with siRNA or pcDNA3.1-Kap1 plasmid transfection, respectively, and then cells were induced for differentiation by 2% horse serum for 4 days, stained for MYHC and DAPI, and fluorescence images were obtained with FV1000 microscope. Scale bar: 100 μm (up) and 50 μm (down). Data are presented as the mean ± SEM (n ≥ 12); *P < 0.05; **P < 0.01. b KAP1 regulates miR-1 and miR-133a expression. The expression of KAP1 in C2C12 cells was manipulated with siRNA or pcDNA3.1-Kap1 plasmid transfection and the efficacy was detected by western blot and quantitative RT-PCR. The influence of KAP1 on miR-1 and miR-133a expression was detected by quantitative RT-PCR. Data are mean ± SEM (n = 4); *P < 0.05; **P < 0.01. c KAP1 regulates C2C12 myoblasts differentiation via miR-133a-related mechanisms. The expression of KAP1 in C2C12 cells was manipulated with siRNA or pcDNA3.1-Kap1 plasmid transfection. Then cell differentiation was induced by 2% horse serum in the presence of miR-133a inhibitor/mimic. The expression levels of KAP1 and MYHC were detected by western blot, and signal intensity was normalized with GAPDH and shown as scatter plot with individual data point graph with significance. Data are presented as the mean ± SEM (n = 3), *P < 0.05; **P < 0.01.