Figure 3.

CCCP treatment causes an imbalance in mitochondrial dynamics in C2C12 myotubes. A, The mitochondrial membrane potential was evaluated as described in the Materials and Methods (left). Intracellular ATP levels were measured in C2C12 myotubes treated with 10 μmol/L CCCP in the presence or absence of 2 mmol/L NAC (right). ATP levels were estimated by using an ATP colorimetric/fluorometric assay kit (Abcam, ab83355) according to the manufacturer's instructions (B) Changes in the mRNA levels of mitochondrial OXPHOS complex subunits after CCCP treatment (C) Measurement of ROS in C2C12 myotubes as described in the Materials and Methods. Scale bars = 50 μm. D, Changes in the expression of proteins related to mitochondrial fusion and fission. E, Immunoblot analyses of proteins related to mitochondrial dynamics in C2C12 myotubes. All results are representative of more than three independent experiments. The data represent the mean ± SEM. P value < .05 obtained by one‐way ANOVA was considered statistically significant