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. 2018 Sep 20;3(18):e99592. doi: 10.1172/jci.insight.99592

Figure 4. Autophagy activity is induced in mouse kidneys after cigarette smoke exposure.

Figure 4

(A) Scheme representing autophagic flux experiment in which leupeptin or bafilomycin A1 inhibits autophagosome degradation, leading to autophagosome accumulation. (B) Mice subjected to RA or CS for 2 months (left panel) or 6 months (right panel) were assayed for autophagic flux in vivo by injection with leupeptin or vehicle (PBS), followed by Western blotting for LC3B expression in kidney tissue after 2 and 6 months of exposure and Beclin-1 expression after 6 months of exposure. Dot plots represent quantitation of Western blots (n = 3 per group, except for 6 months CS + leupeptin exposure, n = 2 per group). (C) HK-2 cells were exposed to CSE in the absence or presence of bafilomycin A1. Representative Western blot of Beclin-1 and LC3B expression to determine autophagic flux in vitro. Dot plots represent quantitation of 3 independent experiments. All data are mean ± SEM.*P < 0.05, **P < 0.01, ***P < 0.001, analyzed by 1-way ANOVA with Bonferroni’s post hoc test.