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. Author manuscript; available in PMC: 2013 Jan 1.
Published in final edited form as: Hepatology. 2011 Dec 6;55(1):222–232. doi: 10.1002/hep.24690

Figure 2. APAP induces autophagy in primary mouse hepatocytes.

Figure 2

Figure 2

Figure 2

(A–B) Ad-GFP-LC3 infected hepatocytes were treated with APAP (5 mM) as indicated (for 3, 6 and 24 hrs) or with different concentrations of APAP (0, 2.5, 5, 10 mM) for 6 hrs and examined by fluorescence microscopy. Scale bar: 20 µm. GFP-LC3 puncta (mean ± SEM) were quantified for each experiment (n=3). At least 30 cells were counted in each individual experiment. *: p<0.01. Total lysates were subjected to immunoblot assay for LC3 and p62 (C). (D) Primary mouse hepatocytes were either treated with saline (panel a) or treated with APAP (5 mM, panels b &c) for 6 hrs and then processed for EM analysis. Panel c is an enlarged photograph from boxed area in panel b. Arrows: autophagosomes, n: nucleus, m: mitochondria; LD: lipid droplets. (E) The number of autophagosomes was quantified from more than 20 different cells (mean ± SD) *: p<0.01. (F) Hepatocytes were treated as in (A). Total cellular lysates were subjected to immunoblot assay for mitochondrial proteins: HSP60 (matrix protein); Cyto c (intermembrane space protein) and Tom20 (outer membrane protein).