AR expression in the skeletal muscle of mice with PCOS and the effects of AR deficiency on the cellular oxidative stress in T-treated C2C12 cells. (A) Representative Western blots and densitometry quantification of AR in skeletal muscle of PCOS and control mice. (B) Representative Western blots of AR expression of T-treated C2C12 cells. Fully differentiated C2C12 cells were treated with T (0, 10 μM) for 12 h. AR expression was measured by Western blot analysis. (C) Representative photomicrographs of the subcellular localization of AR in C2C12 cells. Fully differentiated C2C12 cells were transfected with siRNA-AR or nonsilencing RNA (NC) and then treated with T (0, 10 μM) for 12 h. Red: AR; Blue: DAPI. Scale bar = 25 μm. (D) Representative photomicrographs of intracellular ROS in C2C12 cells. Fully differentiated C2C12 cells were transfected with AR siRNA or non-silencing RNA and then treated with T (0, 10 μM) for 12 h. Intracellular ROS levels were determined by DCFH-DA staining. Sale bar = 100 μm. (E) Relative mRNA levels of NOX4 in T-treated C2C12 cells transfected with siRNA-AR non-silencing RNA (NC). (F) Insulin-signaling assay. (G) Densitometry quantification of pAKT/AKT of the insulin-signaling assay in C2C12 cells. The data are presented as means ± SEM of three independent experiments. *, p < 0.05, **, p < 0.01. T, testosterone; AR, androgen receptor; Ins, insulin.