FIG 3.
Development of DENV3, DENV4, and ZIKV VLPs. (A and B) (Top) Schematic illustrations of VLP expression vectors. (Bottom) Western blot analyses of DENV3 and DENV4 VLPs in culture supernatants. Expression vectors were transfected into 293F cells, and culture supernatants were tested for VLP production on day 4 by using antibodies against DENV1-4 E (A) or DENV4 E (B). The intensity of each band were measured, and the relative intensity was calculated by setting the intensity of the comparator VLPs to 1.0. Images representative of at least 3 replicates are shown. (C) (Left) Schematic illustration of prM-EF108A_ZIKV/JEV. ZIKV E protein aa 404 to 504 were replaced with the corresponding region of JEV, and the F108A mutation was introduced. (Right) Western blot analysis of the ZIKV VLP constructs. Culture supernatants were harvested at 4 days posttransfection, and E proteins were detected by use of a rabbit anti-ZIKV E polyclonal antibody. Images representative of at least 3 independent experiments are shown.