Fig. 4.

Autophagy flux is blocked by ARCSP. a Cells were treated with ARCSP (0–75 μM) for 48 h, and the level of p62 was detected by Western blotting. b p62 mRNA expression after treatment with ARCSP (75 μM). c Cells were treated with ARCSP (0–75 μM) for 48 h, immunolabeling with p62 (594 red) antibodies. DAPI (blue) was used to stain the nuclei, and the cells were photographed under a fluorescence microscope. Scale bar = 25 μm. The histograms showed quantification results of p62 localization which were calculated using ImageJ software. d Cells were transfected with the mRFP-EGFP-LC3 adenovirus, cocultured with ARCSP (75 μM) and Rapa (100 nm) or CQ (20 μM) for 48 h, and analyzed by confocal microscopy. Scale bar = 10 μm. The histograms show the quantification of yellow and red puncta, which was performed using ImageJ software. The data are expressed as the mean ± SD; ^*P < 0.05, ^**P < 0.01, ^***P < 0.001. ns, not significant