Figure 6

Autophagy inhibition resulted in a loop of increasing mitochondrial ROS accumulation under nutrient deprivation. (A) Mitochondrial membrane potential was examined by Rho123 under a fluorescence microscope after EBSS treatment of indicated times. Representative photos of cells were taken a magnification × 400. (B) After indicated treatments for 24 h, Chang liver cells were incubated with MitoSOX Red (red staining) DAPI dye (blue staining). Representative images of cells were taken a magnification × 400 (left). Relative fluorescence intensity of MitoSOX Red per cell for each condition was quantitated in columns. Data were shown as mean ± SEM (n = 3; *: p < 0.05; **: p < 0.01). (C) After indicated treatments for 24 h, oxidative damaged DNA was visualized by immunocytochemistry for 8-OHdG (green) and counter stained with DAPI (blue) at a magnification × 400.