Fig. 5: The increased ROS production in mitochondria and caspase-dependent apoptosis in KLC4-depleted cells.

a KLC4 siRNA-treated R-H460 cells were loaded with the mitochondrion-derived O2– indicator MitoSOX Red and imaged by confocal microscopy. Increased MitoSOX fluorescence indicates the production of mitochondrial ROS in cells treated with KLC4 siRNA. b KLC4 siRNA-treated cells were analyzed by FACS analysis using MitoSOX Red staining in R-H460 and A549 cells. c Mitochondrial ROS analysis of R-H460 cells left untreated or treated with 10 Gy radiation after transfection with KLC4 siRNA, measured 24 h after treatment. d Analysis of Caspase activity in R-H460 cells transfected with KLC4 siRNA after 48 h by ELISA. Data were collected using a Multiskan EX (Thermo) at 405 nm. e Cell death analysis of R-H460 and A549 cells left untreated or treated with 100 µM Z-VAD-FMK (pan-caspase inhibitor) after transfection with KLC4 siRNA, measured 48 h after treatment. f Protein levels of Cleaved PARP and Cleaved caspase3 determined by Western blotting. All data are shown as the mean ± S.D. of at least three independent experiments. The P-values were calculated using unpaired Student’s t-test. *P < 0.05; **P < 0.005; ***P < 0.001