Fig. 5

Effect of lithospermic acid on myogenesis and myotube atrophy in C2C12 cells. A Quantitative HPLC analysis of lithospermic acid and shikonin in GW. B Differentiated C2C12 cells were fixed and stained with total MHC antibody (scale bar, 200 μm). Fusion index was calculated as the average number of nuclei in MHC positive multinucleated cells above total nucleus. C The expression levels of MyoD and MyoG quantified by qRT-PCR in C2C12 myotubes. D After exposure to Dexa with/without LA, myotubes were fixed and stained with α-actinin antibody (scale bar, 200 μm). Myotube diameter was calculated as the average diameter of α-actinin-positive multinucleated myotubes. E The expression levels of Atrogin-1 and MuRF1 quantified by qRT-PCR in C2C12 myotubes. Results are expressed as mean ± SD. One-way ANOVA was used to compare more than two groups, followed by Bonferroni post-hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001 versus the Dexa-treated myotubes