Fig. 4.

Effects of FHL3 knockdown on the differentiation of chicken satellite cells. Chicken satellite cells were transfected with FHL3-siRNA or negative control siRNA and cultured in differentiation medium for the following experiments. a The morphology of cells transfected at 0, 24, 48 and 72 h was observed under the light microscope, and cells at the far right were stained with anti-myosin heavy chain antibody. b After cells differentiation for 72 h, Gemisa staining was used to observe the morphology and the number of nuclei, myotubes in cells. c Fusion index at 72 h of differentiation. The fusion index is calculated as the percentage of total nuclei in cells with three or more nuclei. d The average number of nuclei per muscle tube at 72 h of differentiation. e The relative mRNA expression of Myh7, Myog, MRF4 and Myf5 measured at 0, 24, 48 and 72 h of differentiation. f Western blots detected the protein relative expression levels at 0, 24, 48 and 72 h in Myhc and Myog. β-actin was used as loading control. Data are expressed as means ± sem (N = 3). *P < 0.05; **P < 0.01 versus “Negative siRNA”