C2c12 mouse skeletal muscle cell line

The C2C12 mouse skeletal muscle cell line was purchased from American Type Culture Collection (ATCC, VA, USA). Cell culture reagents and chemicals were from Gibco (Life Technologies, CA, USA). Styrene (STR) and styrene oxide (STO) (Figures 1(a) and 1(b)) and other basic chemical reagents, unless otherwise indicated, were purchased from Sigma-Aldrich (MO, USA). The following antibodies were used in this study: anti-myosin heavy chain (MHC) (Millipore, MA, USA); anti-caspase-9; anti-caspase-8; and anti-caspase-3 (Cell Signaling Technology, MA, USA). Catalase and superoxide dismutase enzyme activity assay kits were obtained from Elabscience (Texas, USA).

The C2C12 mouse skeletal muscle cell line was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) and maintained in high-glucose (25 mM) DMEM (Gibco Corporation, New York, NY, USA) supplemented with 10% FBS, 100 U/mL penicillin, and 100 µg/mL streptomycin at 37 °C in a humidified 5% CO₂ atmosphere. Myoblasts were seeded onto 6-well plates (1.2 × 10⁵ cells/well) for Western blot and qPCR analysis, 12-well plates (6 × 10⁴ cells/well) for immunostaining, and 48-well plates (1 × 10⁴ cells/well) for cell viability measurements. When C2C12 myoblasts reached approximately 90% confluence, the growth medium was replaced with a differentiation medium (high-glucose DMEM containing 2% HS, 100 U/mL penicillin, and 100 µg/mL streptomycin) for 6 days to induce myotube differentiation [32 (link)].