FIG 3.

YM-53601 inhibits HCV production without affecting cell viability. Huh-7.5.1-8 cells were infected with HCV JFH-1 and then treated with increasing concentrations of YM-53601 (0 to 1.5 μM) in serum-free medium. The cells and culture supernatants were harvested on the fifth day postinfection. (A and B) An equal portion of each cell lysate was subjected to immunoblotting for core, NS3, and GAPDH proteins. The results from one representative experiment performed in triplicate are shown. Similar results were obtained in four independent experiments. For panel A, samples were run on two blots and are partially redundant. (C) Total RNA fractions were prepared from cells, and then viral and GAPDH RNAs were quantified by RT-qPCR analysis using specific primers for the core and GAPDH sequences, respectively. The amounts of viral RNA relative to that of GAPDH mRNA are expressed as a percentage of the control value and plotted as a function of the drug concentration. (D) The amount of secreted viral particles in each culture supernatant was determined by ELISA for the core protein and plotted as in panel C. (E) Cell viability was determined by XTT assay and expressed and plotted as in panel C. The data in each graph are means ± standard deviations (SD) for triplicate samples from one representative experiment. Similar results were obtained in two or more independent experiments.