Fig. 4: Robust innate immune responses induced by mutations NS2A from 17D attenuate YFVs in A549 cells.
From: Amino acid changes in two viral proteins drive attenuation of the yellow fever 17D vaccine

a–d, A549 cells were infected with mScarlet reporter YFV (MOI = 0.1). Relative expression by RT–qPCR of IFIT2 (a), IFIT3 (b) and IFNB1 (c), and frequencies of mScarlet-positive cells by flow cytometry (d) at the indicated timepoints. Data are mean ± s.d. from 2 experiments with 3 biological samples (n = 6). Points are coloured by viral genotype and connecting lines are coloured by NS2A allele (black, 17D; grey, Asibi). e–h, A549 cells were infected with mScarlet-17D (e,g) or Asibi-[17D-NS2A] (f,h) at an MOI of 0.1 and treated with DMSO or 2 μM ruxolitinib (Ruxo). Frequencies of mScarlet-positive cells (e,f) and relative expression of IFIT3 (g,h) were evaluated. Data are mean ± s.d. from 3 (e,g) or 2 (f,h) experiments with 3 biological samples (n = 9 or 6). Asibi or Dakar-[17D-Ens], 17D E segment with only non-synonymous mutations compared to Asibi; 17D-[Asibi-Ens], Asibi E segment with only non-synonymous mutations compared to 17D. All other chimaeras include synonymous and non-synonymous mutations in the indicated genomic areas. Two-way ANOVA followed by Tukey’s (a–d) or Šídák (e–h) post hoc test for multiple comparisons (a–d,e–h). P values are only shown for comparisons to Asibi as indicated by colour (a–d) or above selected timepoints (e–h).