Fig. 1: Superior growth kinetics of the rSenegal strain are associated with enhanced efficiency of viral internalization and genome replication in mosquito cells.

A Viral growth kinetics of the rSenegal and rThailand strains were assessed by determining infectious virus titers in the supernatant of ZIKV-infected C6/36 cells (MOI of 0.01) by plaque assay. Data are displayed as mean ± SEM of three biological replicates per group. The efficiency of viral attachment (B), internalization (C), and genome replication (D) was measured in C6/36 cells infected at a MOI of 1. B Viral RNA levels bound to the membrane and in the supernatant were quantified one hour post attachment by incubating cells on ice. Data are displayed as mean ± SEM of six biological replicates per group from two separate experiments. C Internalization efficiency was analyzed by quantifying viral RNA relative to Actin mRNA at 3 h post infection (h.p.i.) following protease E treatment. Internalization was normalized to attached viral RNA levels, in cells infected with the rSenegal strain treated with DMSO or Dynasore (left panel) and in cells infected with the rSenegal or the rThailand strains (right panel). The main bar and vertical error bar represent the mean and SEM, respectively. The left panel comprises six biological replicates per treatment from two separate experiments and the right panel comprises four biological replicates per group. D Genome replication was assessed over 24 h by normalizing ZIKV antigenomic RNA levels to those at 12 h.p.i. (left panel) and genomic RNA levels to those at 0 h.p.i. (right panel). E The virus titer-to-viral genome ratio was analyzed over time by infecting C6/36 cells at an MOI of 0.01 and measuring infectious titers by plaque assay and viral genome copy number by RT-qPCR in supernatants. F Viral decay rate was determined by monitoring infectious titer over time at 28 °C, normalized to titer at 0 h. G The cellular ATP levels of ZIKV-infected C6/36 cells was assessed over time using the CellTiter-Glo assay and normalized to the levels at 0 h.p.i. D–G Data are displayed as mean ± SEM of three biological replicates per group. A–G Statistical significance was determined using two-tailed Student’s t test (*p < 0.05; ns non-significant). Lines and bars are color-coded according to the virus strain. Source data with the exact p values are provided in the Source Data file for Fig. 1.