FIG 6.

Replicative fitness and maturation of evolved DV2 variants. (A and B) Growth kinetics of DV2-WT, DV2-C1, and DV2-V1 in Vero (A) and C6/36 (B) cells. (C) Sequence composition of prM cleavage sites of DV2-WT, DV2-C1, and DV2-V1. (D) Representative Western blot image (bottom) of DV2-WT, DV2-C1, and DV2-V1 viral supernatants blotted with anti-Env and anti-prM antibodies, and quantification (top) of viral maturation (prM/Env) normalized to DV2-WT (lower value = more mature). (E) Competition assay of DV2-WT, DV2-C1, and DV2-V1 at equal FFU ratio in Vero and C6/36 cells at high and low MOIs. Viral genomes were quantified by digital droplet PCR (ddPCR) multiplexed with a general DV2 probe and strain-specific probes. Growth kinetics and maturation of DV2 variants were compared to DV2-WT using two-way ANOVA multiple comparisons.