Ml364

The mouse myoblast cell line C2C12 was obtained from RIKEN Bioresource Center (Tsukuba, Japan). The cells were grown in Dulbecco’s modified Eagle medium (DMEM) containing 4.5 g/L glucose and 10% fetal calf serum (FCS). The cells underwent differentiation into myotubes for 7 days in DMEM containing 4.5 g/L glucose and 2% horse serum. In some experiments, we treated the cells with the USP2‐selective inhibitor ML364 (10 µmol/L) (MedChemExpress, Monmouth Junction, NJ, USA) or vehicle (DMSO, 2 mmol/L) for 0.5, 2, 4, 8, 12 h, or 5 days.

Mouse anti-ErbB2 (clones A-2 and 9G6) antibodies were purchased from Santa Cruz Biotechnology (CA, USA). Goat anti-ErbB2 (AF1129) antibody was obtained from R&D systems. Rabbit anti-USP2 antibody was purchased from Abgent. Mouse anti-GAPDH and anti-β-Actin antibodies were purchased from Proteintech (Wuhan, China). Mouse anti-tubulin antibody was purchased from Sigma. Rabbit anti-EEA1 antibody was purchased from Santa Cruz, and mouse anti-LAMP1 antibody was obtained from BD Medical Technology. Rabbit anti-HER2/ErbB2 (29D8) antibody was purchased from Cell Signaling Technology. Mouse anti-Ubiquitin (P4G7) antibody was purchased from Covance. Secondary (Infrared-labeled) goat anti-mouse and anti-rabbit, and donkey anti-goat antibodies were obtained from LI-COR. Chloroquine and cycloheximide were purchased from Sigma. HSP90 inhibitors 17-AAG, ganetespib, and PU-H71 were purchased from Dalian Meilun Biotechnology Co., Ltd (Dalian, China). ML364 was purchased from Medchem Express.

ML364 was manufactured by MedChemExpress (USA). Ang II was provided by Sigma‐Aldrich (USA). MG132 was a product of Enzo (USA). Anti‐USP2 and anti‐periostin antibodies were purchased from Proteintech (China). Anti‐collagen‐III antibody was manufactured by Abcam (United Kingdom). Anti‐CTGF and anti‐GAPDH antibodies were obtained from Boster (China) and Bioworld (Bioworld Technology, USA), respectively. Antibodies raised against ubiquitin, cyclin D1, P27, β‐catenin, GSK‐3β and P‐ GSK‐3β(Ser9) were provided by Cell Signaling Technology (USA). MTS assay reagents were manufactured by Promega (USA). The Co‐IP assay kit was purchased from Life Technologies (USA). Cell Cycle Detection Kit was manufactured by Keygen (China). The enhanced chemiluminescence (ECL) kit was provided by Santa Cruz Biotechnology (USA). Antibodies were diluted at 1:1000, 1:500 and 1:50 for immunoblot, immunofluorescence and Co‐IP assay, respectively.