FIG. 7.

Inhibition of autophagy at the lysosomal level results in increased exosome production in macrophages and hepatocytes. Macrophages (RAW 264.7) and hepatocytes (Hepal-6) were treated with or without 50 mM of alcohol (EtOH) for 24 hours, and bafilomycin A (100 nM) was added in the last 12 hours of the experiment. After 24 hours, protein levels from macrophages and hepatocytes were analyzed by western blotting for LC3-I and L3-II (A) and p62 (B) using β-actin as a loading control. The densitometry analysis is shown as bar diagrams. The LC3-II and p62 ratio in bafilomycin-treated macrophages and hepatocytes to that of untreated macrophages and hepatocytes was calculated to show reduction in autophagic flux (B). The number of exosomes from macrophages and hepatocytes supernatants after 24 hours was quantified by NTA (C; n = 12). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Abbreviations: Baf, bafilomycin; Con., control.