Figure 4
Expression and localization of DIII-RIgE fusion proteins in insect cells. (a), Western blot analysis. Sf9 were mock-infected (control lane 1), or infected with parental baculovirus vector (control lane 2), recombinant AcMNPV expressing CD16-RIgE (control lane 3), DIII-DENV1-RIgE (lane 4), or DIII-WNVKun-RIgE (lane 5). Cells were harvested at 48 hpi and cell lysates analysed by SDS-PAGE and Western blotting. The 22-23 kDa bands of the envelope DIII-RIgE fusion proteins were detected using monoclonal anti-His6 tag antibody and complementary phosphatase-labeled antibody. (b-d), Confocal IF microscopy. Sf9 cells were infected with AcMNPV expressing DIII-DENV1-RIgE (b), DIII-WNVKun-RIgE (c), or coinfected with AcMNPV-DIII-WNVKun-RIgE and AcMNPV-Pr55GagHIV (d). Cells were harvested at 48 hpi and reacted with monoclonal anti-His6 tag antibody and Alexa Fluor® 488-labeled goat anti-mouse IgG antibody, without cell permeabilization. Nuclei were stained with blue-fluorescent DAPI. Note that the fluorescent signal of the anti-His6 tag localized at the cell surface and outlined the cell contour.