FIG 7.
CM generated from R-848-stimulated THP-1 cells inhibits the Norwalk replicon. THP-1 cells were treated for 12 h with R-848 at 1 or 10 μM to generate CM, which was then tested at a concentration of 50% (vol/vol) against the Norwalk replicon for either 48 or 72 h. Norwalk replicon RNA was quantified by RT-qPCR and normalized to the b-actin gene transcript using the ΔΔCT method. The percent inhibition of replication was calculated relative to mock control (DMSO). ΔCT values for each CM treatment were compared to the values for the mock treatment, and differences were analyzed for significance using an unpaired t test with Welch's correction. The cytotoxic effects of R-848-generated CM were also assessed for each treatment condition. One-way ANOVA statistical analysis was used to measure any significant difference in cytotoxicity between the R-848 CM dosages. All plotted data represent the means ± SEM from two independent experiments performed in triplicate. n.s. (not significant), P > 0.05; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001.