Fig. 4: Introduction of the selected non-synonymous mutations into a ZIKV molecular clone.

ZIKV genome bearing the selected mutations (i.e., single S455L mutation in E, Y87H in NS4B and combined E S455L and NS4B Y87H mutations), and as a reference ZIKV genome without the mutation (i.e., ref no mut), were transfected in Huh7.5.1 cells. a Time-course quantification of infectious viral production at the indicated times post-transfection. Results of 4-to-6 independent experiments; mean ± SD; p-values as indicated in the table above the graph and relative to the reference (ref no mut). b Quantification of the specific infectivity (as in Fig. 1a) in the viral supernatants harvested at 72 h for ZIKV genome mutants or not. Results are the mean ± SD relative to the reference virus set to 1 for each independent experiment; 4-to-6 independent experiments. The p-values indicated in the table correspond to one-by-one comparisons of the condition/mutant displayed below in the graph with conditions/mutants indicated on the left side of the table. c Analysis of the focus size index of the indicated mutated or reference ZIKV determined in Huh7.5.1 and Vero cells, as indicated, for supernatants harvested at 72 h post-transfection. The quantifications are displayed by violin plots, determined as in Fig. 2b. Results of 4 independent experiments; p-values as indicated in the table above the graph. d Time-course analysis of the replication of the E S455L mutant versus reference virus (ref no mut) assessed in activated-TLR3 Huh7.5.1 cells (red lines) as compared to control cells (blue lines). Quantification of the intracellular ZIKV genome levels at the indicated times post-infection at MOI 0.005; 4 independent experiments; mean ± SD. The statistical comparison of intracellular ZIKV GE levels for the same target cells and between mutant and reference virus is indicated in the table at the top of the graphs with indicated p-values and NS: p > 0.05.