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. Author manuscript; available in PMC: 2017 Dec 14.
Published in final edited form as: Toxicol Sci. 2014 Jul 30;142(1):33–44. doi: 10.1093/toxsci/kfu154

FIG. 5.

FIG. 5

Inhibition of autophagy enhances usnic acid-induced apoptosis and decreases cell viability in HepG2 cells. (A) Total cellular protein were extracted at 24 and 48 h after usnic acid treatment, and levels of Bcl-2 family antiapoptosis proteins including Bcl-xl, Bcl-2, and Mcl-1 were detected by Western blotting. GAPDH was used as a loading control. Similar results were obtained from three independent experiments. (B and C) HepG2 cells were pretreated with 10mM 3-MA or 100μM chloroquine for 2 h prior to 24 or 48 h treatment with indicated concentrations of usnic acid. Apoptosis was determined by caspase-3/7 assay (B) and cell viability was assessed by MTT assay (C). The bar graphs show the mean ± SD of three experiments. *p < 0.05, **p < 0.01, and ***p < 0.001 versus treatment with usnic acid alone. (D) Western blotting was performed using antibodies for LC3B, Mcl-1, Bcl-2, and GAPDH.