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 8. Inhibition of RNA transcription, protein synthesis, or proteasome formation does not reverse high Pi-induced myogenin downregulation. (A) Representative whole-cell lysate immunoblots for Nrf2, p62, myogenin, and LC3B. Differentiated C2C12 cells were pretreated for 2 h with Act D (10 nM), CHX (5 μg/ml), or MG-132 (0.1 μM) and then exposed for 24 h to the indicated Pi concentrations. (B) Immunoblot analysis of Nrf2, p62, myogenin, LC3B, MYH, and MLC-2v content in protein aggregates from differentiated C2C12 cells treated for 24 h with the indicated Pi concentrations. (C) Immunoblot analysis of Nrf2, p62 and myogenin expression in whole cell lysates from differentiated C2C12 cells treated for the indicated times with CHX or MG-132, with or without 4 mM Pi.