CRVP379 interacts with mosquito prohibitin during DENV infection.

A. Prohibitin is required for DENV infection of Aedes aegypti. Mosquitoes were intra-thoracically injected with either dsRNA against the coding region of prohibitin (Prohdsrna) or dsRNA against GFP (gfpdsrna) as control. At 4 dpmi, mosquitoes were infected with DENV through blood feeding. At 4 dpi, midgut tissues were dissected and individually analyzed for gene expression with qRT-PCR analysis. Each data point represents one mosquito midgut. P<0.01. B. Aag2 cells were transfected with His-tagged CRVP379 and/or infected with DENV. At 48h post-infection, antibodies against the His tag were used to pull the His-tagged CRVP379 protein out of cell lysates. The precipitated solution was run on SDS-PAGE gel and Western blot analysis was performed using antibody against prohibitin. C. CRVP379 and prohibitin co-localize in mosquito cells during DENV infection. Aag2 cells were transfected with His-tagged CRVP379 and infected with DENV 48h post-transfection. Cells were fixed in 4% paraformaldehyde 24h post-infection and stained with antibodies against the His-tag (green) and against prohibitin (red). DAPI was used to localize the nucleus. Representative images are shown.