FIG 4.
VCP/p97 activity is required for flavivirus infection. (A) Nluc expression at 18 h postinfection with YFV-17D/Nluc (MOI of 0.3) of cells treated with the indicated compounds: Pyr-41 (50 μM), DBeQ (10 μM), NMS-873 (300 nM), or CHX (100 μg/ml). (B) Nluc expression at 5 h posttransfection with 0.2 ng YFV-17D/Nluc RNA in cells treated with the indicated compounds, as described above. (C) Infection of BHK cells with WNVKUN or ZIKV (each at an MOI of 1.0) in cells treated with the indicated compound, as described above. Cells were fixed at 24 h postinfection, stained for NS1 expression, and counted by flow cytometry, as detailed in Materials and Methods. (D) Nluc expression at 6 h after pretreating C6/36 mosquito cells for 1 h with the indicated compounds, followed by infection with YFΔSK/Nluc (MOI of 0.3) or transfection with 20 ng of YFΔSK/Nluc RNA (as indicated). The experiments depicted in panels A and D were performed three times, each in triplicate. The experiments depicted in panels B and C were performed twice in parallel, each in triplicate. Statistical significance was calculated by using one-way ANOVA (****, P < 0.0001; ***, P < 0.001; **, P < 0.01; *, P < 0.05; ns, not significant).