Zln005

The mouse C2C12 myoblasts (American Type Culture Collection, Manassas, United States) were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Thermofisher, Massachusetts, United States) supplemented with 10% fetal bovine serum (Thermofisher, Massachusetts, United States), 2 mM glutamine, 100 IU/ml penicillin, and 100 mg/ml streptomycin (Merk Millipore, Germany) at 37°C with saturated humidity and 5% carbon dioxide. Trypsin-EDTA was used to detach cells from tissue culture dishes. The dissociated C2C12 myoblasts were subcultured in a new dish. When the myoblasts grew to 70–80% confluence, the culture medium was switched to differentiation medium consisting of DMEM containing 2% horse serum (Thermofisher, Massachusetts, United States) for 5 days to induce myoblasts to differentiate into myotubes. In this study, C2C12 myotubes were treated with drugs, including PGC-1α activator ZLN005 (MedChemExpress, New Jersery, United States), Human/Murine/Rat Irisin (Pepro Tech, New Jersery, United States), Recombinant Human/Murine/Rat Myostatin (Pepro Tech, New Jersery, United States), activin-like kinase (ALK) 4/5 inhibitor TEW-7197 (Selleck, Shanghai, China), Smad3 inhibitor SIS3 (Selleck, Shanghai, China), ERK inhibitor U0126 (Selleck, Shanghai, China), and a vehicle (dimethylsufoxide, DMSO).

The human ovarian cancer cell lines A2780 and SKOV3 were obtained from the Shanghai Cell Bank of Chinese Academy of Sciences (Shanghai, China). Both cell lines were cultured in RPMI-1640 medium (Gibco, Carlsbad, CA, USA). Cisplatin (CDDP) and 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Epoxomicin (Epox), GSK3β inhibitor (CHIR99021), and the PGC1α activator ZLN005 were purchased from MedChemExpress (Monmouth Junction, NJ, USA). Transfections were performed using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA). Anti-Keap1 and anti-lamin B antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-p38, anti-GSK3β, anti-phospho-p38, anti-phospho-GSK3β, and anti-AKT antibodies were acquired from Cell Signaling Technology (Danvers, MA, USA). Anti-Nrf2, anti-Bcl-2, anti-Bax, anti-cleaved caspase 3, and anti-β-actin antibodies were procured from Proteintech (Chicago, IL, USA). Total OXPHOS Human WB Antibody Cocktail and anti-PGC1α antibodies were purchased from Abcam (Cambridge, MA, USA).