Fig. 3

TMZ-POH induces mitophagosome accumulation. a Total protein, mitochondrial protein and cytoplasmic protein extracted from A549 and SPC-A1 cells were subjected to western blot analysis for demonstrating LC3B expression. ACTB acts as control for total and cytoplasmic protein, HSP60 acts as control for mitochondrial protein; (b) A549 cells were either starved or treated with TMZ-POH, and stained with anti-LC3B and anti-TOM20 antibodies and imaged by confocal microscopy; Statistical analysis of the colocalization coefficient of LC3B and TOM20 using Fiji Image J program; (c) A549 cells were treated with TMZ-POH or not, and stained with anti-Parkin and anti-TOM20 antibodies and imaged by confocal microscopy; Statistical analysis of the colocalization coefficient of Parkin and TOM20 using Fiji Image J program; The results shown are means ±SD, **p < 0.005, ***p < 0.001