Fig. 2

CISD2 knockdown promotes the activation of autophagy. A Representative images of the LC3 punches formation by confocal microscope in HT-1080 cells transfected with LC3-GFP adenovirus with or without Erastin (7.5 µM) treatment for 2 h. Scale bar: 10 μm. B Detection of the activity of lysosome by Magic Red cathepsin B staining and confocal microscopy. The intensity of red fluorescence was positively correlated with the lysosomal activity. Scale bar: 10 μm. C, D Statistical graph of LC3-GFP and cathepsin B staining, respectively (n = 3). E Western blot analysis of the expression of proteins associated with autophagy; GAPDH was used as loading control. F Quantitative analysis of LC3 expression; ^★★P < 0.01 between the indicated groups