2.5. JC-1 Staining

The mitochondrial membrane potential was determined in the C2C12 myotube cells with JC-1 kits (Beyotime, Shanghai, China). After PBS washing, the cells were stained with JC-1 for 20 min at 37 °C. The fluorescences were detected using a fluorescence microscope. Then, the regions were randomly selected from each group and the relative fluorescence intensity of cells were measured by the software Image J.