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. 2011 Oct 5;286(48):41563–41577. doi: 10.1074/jbc.M111.280073

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© 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 7. — Reduced levels of HRD1 induce autophagy in ATZ-expressing Hepa 1-6 cells. A, Hepa 1-6 cells were transfected with LC3-pEGFP-C1, AAT-myc-pcDNA3.1, and either control siRNA (csiRNA) or HRD1 siRNA (panel i) or LC3-pEGFP-C1, ATZ-myc-pcDNA3.1, and either control siRNA or HRD1 siRNA (panel ii). Where indicated, autophagy was induced by starvation (st) 48 h after transfection. Cells were fixed, and primary staining was done using the mouse monoclonal antibody against Myc to visualize ATZ. Secondary staining was carried out using Cy3-conjugated anti-mouse antibodies. Bar, 25 μm. B, quantification of the experiment shown in A (AAT + control siRNA, n = 29; AAT + control siRNA + starvation, n = 8; AAT + HRD1 siRNA, n = 23; ATZ + control siRNA, n = 80; ATZ + control siRNA + starvation, n = 40; ATZ + HRD1 siRNA, n = 26). Statistical significance is indicated by *** (p ≤ 0.0001) and ns (p > 0.05). Error bars = standard deviations.