Fig. 4

1700113A16Rik was required for the differentiation of MuSCs. A Immunofluorescent staining of MyHC in the control group (siNC) and 1700113A16Rik knockdown groups (si1700113A16Rik-1 and si1700113A16Rik-2). MuSCs were transfected by two pieces of siRNA against 1700113A16Rik or scramble RNA one day prior to differentiation and then differentiated for 3 days. The differentiated MuSCs were stained with anti-MyHC. Green indicated MyHC; DAPI indicated nuclear staining; merge indicated the merged images of green and blue. (Scale bars, 500 μm). B Statistical analysis of the number of MyHC positive cells, the myotube length and fusion index of myotubes described in A. Error bars indicated standard deviation and were based on 3 independent experiments. * indicated p < 0.05, *** indicated p < 0.001. C The relative RNA level of 1700113A16Rik and Myh1 in the differentiated MuSCs described in A. RT-qPCR assays were performed with RNA extracted from the differentiated MuSCs in the control group and the 1700113A16Rik knockdown group. Error bars indicated standard deviation and were based on 3 independent experiments. * indicated p < 0.05, ** indicated p < 0.01. D The protein level of MyHC in the differentiated MuSCs described in A. Western blot was performed from the differentiated MuSCs in the control group and the 1700113A16Rik knockdown group. The numbers below each panel indicated the relative signal intensity. E Immunofluorescent staining of MyHC. MuSCs were infected by control adenovirus and adenovirus encoding 1700113A16Rik and then differentiated for 3 days. The differentiated cells were subjected for immunofluorescent staining with MyHC antibody. Red indicated MyHC; Blue indicated DAPI; Merge indicated the merge of red and blue images. (Scale Bars, 200 μm). F Statistical analysis of the number of MyHC positive cells, the myotube length and fusion index of myotubes described in E. Error bars indicated standard deviation and were based on 3 independent experiments. * indicated p < 0.05, ** indicated p < 0.01