Anti cox1

The total proteins were extracted from proliferating or differentiating MuSCs (1 × 10⁶ cells per well) transfected with an AgomiR-145-3p, miR-145-3p mimics, LNAantimiR-145-3p, or miR-145-3p inhibitor with their respective controls, using protein lysis radioimmunoprecipitation assay (RIPA) buffer containing 1 mM PMSF (Solarbio, Beijing, China) 72 h post-transfection. Subsequently, proteins in the supernatant were separated by SDS-PAGE, and then transferred to 0.2 mm polyvinylidene fluoride (PDVF) membranes and sealed with 5% skim milk for 2 h at room temperature. After, the membranes were incubated with primary antibodies specific for anti-COX1, anti-MYBL1, anti-PCNA, anti-Pax7, anti-beta-tubulin, anti-Myomaker, anti-MyoD, anti-MyHC, and anti-MyoG (ABclonal Biotechnology Co., Ltd., Hubei, China) at 4 °C overnight. The PVDF membranes were washed with Tris saline with Tween (TBST) buffer 3 times and then incubated with horseradish peroxidase (HRP)-conjugated secondary antibodies for 2.5 h at room temperature. The enhanced chemiluminescence luminous fluid (ECL) was applied for color development.