Fig. 7.

Effect of M-cadherin RNAi and GSK-3β inhibition on myogenic differentiation of C2C12 myoblasts. C2C12 cells were grown on coverslips, transfected with M-cadherin-targeted (M-) or non-targeted scrambled siRNA (SiCON) for 36 hours then incubated with or without TDZD-8 for 12 hours. The cells were then cultured in differentiation medium for 48 hours. (A) Representative confocal images of C2C12 myoblasts after treatment with a combination of M-cadherin RNAi (M-) or non-targeted scrambled siRNA transfection (SiCON) and TDZD-8 or DMSO as a vehicle control. The cells were cultured in differentiation medium for 48 hours after the co-treatments. MyHC, red; DAPI, blue; TUNEL, green. Scale bar: 100 μm. (B,C) Data from three independent experiments as mean ± s.e.m. *P<0.05 vs SiCON/DMSO; †P<0.05 vs M-/DMSO. The myoblast fusion index was calculated for cells after each treatment described in Fig. 7A, as the ratio of the number of DAPI-positive nuclei located in the MyHC-positive myotubes (i.e., fused myoblasts) divided by the total number of nuclei in the same field (B). The apoptotic index was calculated as the percentage of total nuclei that were TUNEL positive (C).