Silencing select virally-up-regulated genes reduces DENV infection in mosquito cells.

A. The mosquito genes listed in S1 Fig were knocked down in Aag2 cells using RNAi and the effects on DENV infection were analyzed. The genes that reduced infection below 60% of control are shown. Aag2 cells were infected with DENV (MOI of 1.0) 72 h post-knockdown and analyzed for infection by qRT-PCR 24h post-infection. Data is displayed as percent control infection (using scrambled siRNA). Both DENV infection and qRT-PCR analysis were done in triplicate, data is pooled and error bars indicate standard deviation. B. DENV infection increases CRVP379 in Aag2 cells over time. Aag2 cells were infected with DENV (MOI of 1.0) and infection was measured using qRT-PCR analysis at the timepoints indicated. P<0.01. C. Expression of CRVP379 during RNAi knockdown. CRVP379 siRNA was transfected into Aag2 cells and gene expression was analyzed by qRT-PCR. Samples were taken at 24, 48 and 72h post-knockdown. Expression after transfection of GFP control siRNA is also indicated. D. Reduction of CRVP379 reduces DENV infection over time. Either siRNA against CRVP379 or GFP was transfected into Aag2 cells and cells were infected with DENV (MOI of 1.0) at 72 h post-knockdown. Cells were analyzed for infection by qRT-PCR at the timepoints indicated. E. DENV infection increases CRVP379 expression during siRNA knockdown. Either siRNA against CRVP379 or GFP was transfected into Aag2 cells and cells were infected with DENV at 72 h post-knockdown. Gene expression was analyzed by qRT-PCR at the timepoints indicated. Data is expressed as the fold change in CRVP379 expression in cells with GFP siRNA versus cells with CRVP379 siRNA during DENV infection. B-E. Data is pooled from 6 separate experiments, error bars indicate standard deviation.