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. Author manuscript; available in PMC: 2014 Nov 20.
Published in final edited form as: Nat Commun. 2014 May 20;5:3877. doi: 10.1038/ncomms4877

Figure 5. prM H98A mutation inhibits DENV maturation and infection.

Figure 5

a. Test of infectivity. RNAs derived from the WT and prM H98A DENV1 WP infectious clones were electroporated into 293T cells. Progeny virus was collected four days post-electroporation and titered on BHK-21 cells. Data are the mean and SD of three independent experiments, with WT titers normalized to 105 IC/ml. b. Test of prM processing. RNAs derived from the WT and prM H98A DENV1 WP infectious clones were electroporated into 293T cells. Cells were radiolabeled with 35S methione/cysteine for 4 days at 29 °C. Virus in the labeling medium was pelleted by ultracentrifugation, immunoprecipitated with mouse anti-DENV2 serum and analyzed by SDS-PAGE and phosphorimaging (“virus”). The cell lysates were analyzed by SDS-PAGE and western blot using mAbs to E protein and β-actin (“Cell”). Positions of molecular weight markers are shown on the left. Data are a representative example of three independent experiments. Full scans of the phorphorimages and blots are in Supplementary Fig. 4.