FIG 7.

Effect of DPM1 depletion on DENV viral protein glycosylation. Control, DPM1^KO, and STT3A^KO 293T cells and DPM1^KO cells complemented with the WT or catalytically dead mutants of DPM1 were cotransfected with DENV C-prM-E and NS2B-NS3 plasmids (1:1 ratio) (A) or transfected with DENV hemagglutinin (HA)-tagged NS1 plasmid (B). Cell lysates were subjected to immunoblot analysis with anti-capsid, anti-prM, anti-E, and anti-HA antibodies. Data are representative of results from three independent experiments. (C) Control and DPM1^KO 293T cells were cotransfected with DENV C-prM-E and NS2B-NS3 plasmids or HA-NS1 plasmid. Twenty micrograms of total proteins was subjected to deglycosylation with either PNGase F or endo H (25 kU ml⁻¹) for 1 h at 37°C and analyzed by immunoblotting with anti-prM, anti-E, and anti-HA antibodies. As a positive control for deglycosylation, cells were also transfected in the continuous presence of tunicamycin (Tun; 5 μg ml⁻¹). For all panels, blots are representative of those from three independent experiments. NT, nontreated.