Figure 6
From: PGC-1α buffers ROS-mediated removal of mitochondria during myogenesis

Downregulation of FOXO1 enables mitophagy induction. C2C12 cells were transfected with scramble (scr), PGC-1α and/or FOXO1 siRNA (PGC-1α(−), FOXO1(−) and PGC-1α/FOXO1(−)) and differentiated in DM for the indicated days. (a) Twenty micrograms of total proteins were loaded for western blot analysis of FOXO1, PGC-1α, SQSTM1 and PINK1. TUBB was used as the loading control. Numbers indicate the density of immunoreactive bands calculated using the Software Quantity one (Bio-Rad) and reported as the ratio of SQSTM1/TUBB and PINK1/TUBB. (b) Twenty micrograms of total proteins were derivatized with DNP and carbonylation was detected by western blot with DNP antibody. TUBB was used as the loading control. (c) Twenty micrograms of total proteins were loaded for western blot analysis of FOXO1, PGC-1α, PINK1 and BNIP3. TUBB was used as the loading control. Numbers indicate the density of immunoreactive bands calculated using the Software Quantity one (Bio-Rad) and reported as the ratio of proteins/TUBB. Immunoblots reported in the figures are representative of at least four experiments that gave similar results