Research Paper Volume 12, Issue 21 pp 21446—21468

High phosphate induces skeletal muscle atrophy and suppresses myogenic differentiation by increasing oxidative stress and activating Nrf2 signaling

Figure 3. High Pi impairs mitochondrial function in differentiated C2C12 cells. Mitochondrial function was assayed by determining MMP, OCR, and ECAR in 3-day-differentiated C2C12 cells treated with the indicated concentrations of Pi for 24 h. (A) Flow cytometric analysis of MMP (JC-1 staining). Cells exposed to H2O2 (10 mM) for 1 h served as positive control. (B) OCR (pmol/min) and (C) ECAR (mpH/min) were measured in proliferating (10% FBS) and differentiated (2% HS) C2C12 cells using a Seahorse XF24 analyzer. (D) Comparison of basal OCR between proliferating and differentiated C2C12 cells. (E) OCR and (F) ECAR measurements in 3-day-differentiated C2C12 cells treated with the indicated concentrations of Pi for 24 h. (G) OCR/ECAR ratios during basal and maximal respiration in differentiated C2C12 cells treated with the indicated concentrations of Pi. Data are presented as means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, compared to control (vehicle).