Fig. 2: Structural genes enhance viral internalization, and non-structural genes improve genome replication of rSenegal strain relative to rThailand strain in mosquito cells.

A Schematic representation of the first set of chimeric viruses. B Plaque morphologies of chimeric viruses on Vero E6 cell monolayers. Plaque diameters were measured using ImageJ software for quantification. Data are displayed as mean ± SEM of six or seven biological replicates per group. C Viral growth kinetics of chimeric viruses with rSenegal (left panel) or rThailand (right panel) backbones were determined by measuring infectious titers from supernatants of ZIKV-infected C6/36 cells (MOI = 0.01) by plaque assay. The efficiency of viral internalization (D) and genome replication (E) was analyzed in C6/36 cells infected with ZIKV at an MOI of 1. Viral RNA levels were assessed by calculating the ratio of viral RNA to Actin expression at 3 h.p.i. for internalization (D) and over 24 h for genome replication (E) following protease E treatment. Internalization efficiency (D) was determined by normalizing viral RNA at 3 h.p.i. to levels of initially attached viruses. Genome replication (E) was evaluated by normalizing ZIKV antigenomic RNA levels at each time point to 12 h.p.i. (left panel) and genomic RNA levels to baseline levels at 0 h.p.i. (right panel). F The cellular ATP levels following ZIKV infection was assessed over time using the CellTiter-Glo assay and normalized to the levels at 0 h.p.i. C–F Data are presented as mean ± SEM of three biological replicates per group. B–F Statistical analysis was performed using one-way ANOVA with Dunnett’s test (*p < 0.05; **p < 0.001; ns non-significant). Lines and bars are color-coded to represent the different chimeric viruses, and the parental strains are depicted with thicker lines. Source data with the exact p values are provided in the Source Data file for Fig. 2.