Counteraction of HCV-Induced Oxidative Stress Concurs to Establish Chronic Infection in Liver Cell Cultures

Figure 1

Generation of an in vitro model of acute and chronic HCV infection. (a) Schematic representation of the strategy to obtain long-term infected culture (LTIC). The fluorescence microscopy pictures were obtained from one representative experiment out of three performed. Cells were labelled with anti-hepatitis C virus core 1b protein followed by Alexa Fluor 488-conjugated secondary antibody (green); the nuclei were stained with DAPI (blue), 40× objective; (b) virus titration by qRT-PCR at different times of infection. (c) Trypan blue dye exclusion analysis of cell viability in uninfected (CTR) and HCV-infected Huh7.5 cells at different days postinfection. from three independent experiments are shown. ; . (d) Biparametric flow cytometry analysis of apoptosis at different time points after HCV infection. In the left panel, a representative dot blot of apoptosis detection is shown relative to the samples obtained from 14 days p.i. Numbers represent the percentage of apoptotic cells either annexin V/PI double-positive (upper right quadrant) or annexin V single-positive (low right quadrant). In the right panel, the bar graphs show the of the percentage of annexin V/PI-positive cells expressed as percentage of variation vs. control uninfected cells. The mean values were obtained from four independent experiments. vs. CTR uninfected cells.

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