Figure 1.

A: Hep-G2 cells, stably transfected with full length HCV (Hep-394) or empty vector (Hep-SWX), were plated in 96-well plates and incubated with varying concentrations of sorafenib. Cell viability was assessed by the CellTiter 96 AQueous assay kit after 72 hours of exposure to drug or diluent control. IC50 values were calculated after curve-fitting using the XLfit Software. Error bars indicate SEM (n=6). B: Cells were plated in 35-mm dishes and stained with DAPI at the indicated time points. Cells with morphological changes of apoptosis were counted using a fluorescence microscope and the number of apoptotic cells expressed as a percentage of the total cells. Error bars indicate SEM. * p = 0.03 (n=3). C: Hep-SWX and Hep-394 were incubated in serum free medium and caspase 3/7 activation was assessed after 72 hours using a luminometric assay. Error bars indicate SEM. * p = 0.003 (n=3). D: Cell lysates from Hep-SWX and Hep-394 cells were obtained and evaluated for the expression of PARP by western blotting. Representative blots are shown along with the average and SEM of the expression of cleaved PARP in Hep-394 cells relative to the expression in Hep-SWX from three separate experiments. p: 0.002. E: Hep-SWX and Hep-394 cells were incubated in serum-free medium with sorafenib (2μM) or DMSO for 72 hours and the apoptotic cells quantitated by TUNEL assay and flow cytometry. Bars represent mean and SEM of 4 independent experiments. * p< 0.05 relative to Hep-SWX for each condition. F: The expression of Mcl-1 was analyzed by immunoblot analysis. Representative blots are shown along with quantitative data representing the average and SEM from three separate experiments. The relative expression of Mcl-1 to actin is expressed relative to that in Hep-SWX cells. p < 0.001.