Figure 2. Inhibition of HCV replication and infectious virus production.

Huh-7.5 cells were infected with the HJ3-5 virus at a multiplicity of infection (MOI) of 1, and 72 h later, DMSO or Peretinoin was added at the indicated concentrations. The medium was replaced with fresh medium every 24 h until 72 h. (A) At 72 h after adding Peretinoin, total cellular RNA was extracted, and the amount of HCV RNA and 18S rRNA was quantitated by RTD-PCR. Relative HCV RNA abundance normalised to the amount of 18S rRNA is presented as fold change ± SD compared to DMSO-treated cells from 3 independent experiments. (B) At 72 h after Peretinoin treatment, the cell lysates were collected and subjected to western blot analysis using anti-core protein and anti-β-actin antibodies. Full-length blots/gels are presented in Supplementary Fig. S2 online. (C) The medium was collected at 72 h after Peretinoin treatment, and immediately, naïve Huh-7.5 cells were infected with serially diluted medium. At 72 h after infection, the infectious virus titre of HCVcc from Peretinoin-treated cells was determined by an FFU assay. Data shown here represent the mean FFUs/mL ± SD from 2 independent experiments.