Fig. 2.

Inhibition of autophagy suppressed replication of HCV replicon. a Cells were treated with 3-methyladenine (3-MA) (10 mM) or a mixture of E64d (1 μg/ml) and pepstatin A (Pep) (1 μg/ml) for 18 h, the levels of replication of HCV replicon were assessed by luciferase assay. b Cells were treated with 3-methyladenine (10 mM), mixture of E64d (1 μg/ml) and pepstatin A (1 μg/ml) for 18 h. Cell proliferation reagent WST-1 was added to each well, and the cells were incubated for 1 more hour at 37°C. The absorbance was measured against a background control by microplates reader at 450 nm. The reference wavelength was 650 nm. c A combination of four chemically synthesized siRNA duplex molecules targeted to the human ATG5, 7, LC-3α, LC-3β mRNA sequence was transiently transfected into Huh7/Rep-Feo cells using a transfection reagent. siRNA targeted to enhanced green fluorescence protein was used as the control. Forty-eight hours after transfection, levels of HCV replication were analyzed by luciferase assay