FIG 5.

Mutations in NS4A affecting the interaction between the NS4A-2K-4B precursor and NS1. (A) Schematic representation of the experimental approach. Plasmids encoding the NS4A-2K-4B precursor and containing mutations in NS4A were transfected into Huh7/Lunet-T7 cells stably expressing HA-tagged DENV NS1. Cell extracts were prepared 16 h posttransfection and used for HA-specific immunoprecipitation. Captured immunocomplexes were analyzed by SDS-PAGE and Western blotting. (B) Western blot analysis of captured immunocomplexes. A representative experiment of three independent experiments is shown. (C) Quantification of coimmunoprecipitation experiments showing the ratio between HA-NS1 and the NS4A-2K-4B precursor. Values were normalized against the wild-type (WT) sample that was set to 1. Data are the mean and SD from three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001, as determined by two-tailed t test.