Fig. 6.

Protein extracts from S. macrurus blastema (SmBE) inhibits differentiation of C2C12 myoblast cells. Myoblasts were induced to differentiate for 5-6 days in the presence of differentiation medium (DM) only (Control), adult muscle extract (AM), or SmBE (1-week and 2-week) at concentrations of 0.025 mg/ml (0.025) and 0.05 mg/ml (0.05). A: Cells were fixed and immunostained using MF20 (green in merged images). Cell nuclei were stained with Hoechst 33342 (blue in merged images). B: Quantitative analysis for the effects of SmBE on muscle differentiation. a: Effect of SmBE on MHC expression. b: Effect of SmBE on myotube formation. c: Effect of SmBE on cell fusion measured by number of nuclei per myotube. Data are the means ± SD. Student t-test was used to compare the effect of extract with control. Asterisk (*) indicates a difference at P < 0.05. C: Western blot analyses of C2C12 myoblasts cultured in DM for 5-6 days in the presence of different protein extracts. Total protein (50 μg) extracted from myoblasts were resolved by 4-15% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to a polyvinylidene difluoride (PVDF) membrane, and probed with MF20. The membrane was also probed with monoclonal β-actin antibody to demonstrate equal loading of protein.