FIG 6.

The DPMS complex is required for optimal viral RNA replication. (A) Control, DPM3^KO, and DPM3^KO 293T cells complemented with DPM3 were inoculated with DENV 11681 (MOI of 10). At 4 and 24 hpi, cells were treated with trypsin to remove cell surface-bound virus and viral RNA was quantified by RT-qPCR. Data are means ± SD from two independent experiments performed in triplicate. Significance was calculated using a one-way ANOVA with Tukey’s multiple-comparison test. ***, P < 0.001; ****, P < 0.0001). (B) Control, DPM1^KO, DPM3^KO, and STT3A^KO 293T cells were transfected with in vitro-transcribed subgenomic DENV2 RNA expressing the Renilla luciferase. Rluc activity was monitored at the indicated time points. Data are means ± SD from two independent experiments performed in quadruplicate. Significance was calculated using a two-way ANOVA with Dunett’s multiple-comparison test. **, P < 0.01; ****, P < 0.0001. (C) Control, DPM1^KO, and DPM1^KO 293T cells complemented with DPM1 were inoculated with DENV2 16681 (MOI of 100). Left, representative images of the cells stained with anti-dsRNA MAb J2 at 24 hpi. Right, quantification of the number of foci per cell using Icy software. Data are means ± SD from a representative experiment with n = 50 independent cells. Significance was calculated using a one-way ANOVA with Dunnett’s multiple-comparison test. ****, P < 0.0001.