Fig 5. Effect of TAK-242 on LPS-induced inhibition of myogenesis.

(A) May–Grünwald–Giemsa staining of C2C12 cells cultured for 144 h in DM alone, DM plus LPS (1 μg/mL), or DM plus LPS (1 μg/mL) and TAK-242 (1 μM). Representative images are shown. Scale bar = 100 μm. Arrows indicate differentiated myotubes. (B) TAK-242 partially reversed the LPS-induced decrease in myogenic index. Cells were treated as described in (A). Data are the mean ± SEM of 10 independent experiments per treatment group, each examining 5 randomly selected fields (total 50 fields). (C and D) TAK-242 partially ameliorated the LPS-induced myotube atrophy. Cells were treated as described in (A), and the distribution of myotube widths (C) and mean myotube widths (D) were calculated for each treatment group. Data are the mean ± SEM of 10–12 independent experiments, each examining 5 randomly selected fields (total 194–296 myotubes from 50–60 fields per treatment group). (E) Representative western blot probed with antibodies to MyHC II or β-tubulin (internal standard). Cells were treated as described in (A). (F) Quantification of the data presented in (E). Data are the mean ± SEM of 12–13 independent experiments. ***p < 0.001, ###p < 0.001, #p < 0.05 by one-way ANOVA followed by Tukey’s honest significant difference test.