Fig. 3.

TCC inhibition induced ERS and increased apoptosis in C2C12 myotubes. C2C12 myotubes were treated with different concentrations of NNC‐55 after 4 days of differentiation according to the description in Fig. 1. (A) The intracellular ERS analysis by ThT assay under a fluorescence microscope and mean ThT fluorescence intensities were presented (C); ThT stained green and DAPI stained blue; scale bar, 20µm; (B) mRNA expression of ERS‐involved genes including GRP78 and CHOP by qRT‐PCR analysis in C2C12 myotubes. (D) Protein levels of GRP78, CHOP, cleaved‐caspase 3, and cleaved‐caspase 9 were analyzed by western blot and quantitation of proteins (E). These data are presented as the (mean ± SD) for three independent experiments. **P < 0.01, ***P < 0.001, ****P < 0.0001 vs Control, respectively. Data were analyzed using one‐way ANOVA and Dunnett's test.