FIG 2.

SB258585 inhibits HCV infection, modulating PKA in a 5-HT6-independent manner. (A) Huh-7 cells were treated with SB258585 at different concentrations for 2 h prior to, during, or after HCVcc JFH1 incubation, following the schematized kinetics. Infection at 30 h postinfection was quantified by immunofluorescence assay. (B) Huh-7 cells were treated with the drug for 2 h, followed by 28 h of rest. An MTS assay was performed in order to evaluate cell toxicity. (C) Quantification of 5-HT6 mRNA levels in 17 tissues from human biopsy specimens by qRT-PCR. a.u., arbitrary units. (D) Huh-7 cells were treated for 2 h with DMSO, H89 (10 μM), SB258585 (100 μM), or SB399885 (100 μM). A representative Western blot (n = 3) and relative quantification of the total phosphorylation of PKA substrates normalized to the loading control (β-tubulin) are presented. Results are presented as means ± SEM (n = 3) in panels A, B, and D. One-way (B and D) or two-way (A) analysis of variance (ANOVA) followed by the Dunnett or Bonferroni posttest was performed for statistical analysis. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.001.