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. 2021 Jun 10;12:650055. doi: 10.3389/fphys.2021.650055

FIGURE 3.

FIGURE 3

Nicotine caused mitophagy impairment through decreased the CTSL enzyme activity via ROS mediated p38/JNK pathway. (A) Nicotine decreased the activity of CTSL. (B) Torin1 alleviated the activity of CTSL damaged by nicotine. (C) Immunofluorescence co-localization analysis was used to detect the effect of Torin1 after nicotine treatment. Representative of n = 3 experiments. (Scale bar = 20 μm). (D) Western blotting was used to detect the expression of p62 and LC3. (E) Western blotting was used to detect the activation of mTOR pathway. (F,G) Mito-ROS was detected by mito-SOX (Scale bar = 20μm). (H) CTSL activity was restored with NAC pretreatment (I) Mitophagic flux impairment was rescued by NAC, SP100625 and SB203580. Representative of n = 3 experiments. (Scale bar = 20 μm). (J) Western blotting was used to detect the expression of p62 and LC3. (K) Image J was used to quantification (****p < 0.0001. ***p < 0.001. **p < 0.01. *p < 0.05, n = 3).