Fig. 6

RIG-I and MDA5 are essential for macrophage polarization in vivo during WNV infection. WT or RIG-I/MDA5 DKO mice were infected with 100 pfu of WNV for 6 days. Spleens, brains, and IP lavage were harvested and analyzed. a Flow cytometry gating strategy for identification of CD11b⁺ F4/80⁺ cells, M1, and M2 populations. Representative histograms for STAT1pY701 and STAT6pY641 shown on right from three infected spleen samples and a representative mock from each genotype. b Absolute number of CD11b⁺ F4/80⁺ cells (left column), M1 (STAT1pY701⁺) cells (middle) and M2 (STAT6pY641⁺) cells (right column) from the IP lavage (top), spleen (center), or brain (bottom) from WT mice (blue) and RIG-I/MDA5 DKO (red). c Frequency of CD11b⁺ F4/80⁺ cells (left column), M1 (STAT1pY701⁺) cells (middle) and M2 (STAT6pY641⁺) cells (right column) as measured by flow cytometry from the IP lavage (top), spleen (center), or brain (bottom) from WT mice (blue) and RIG-I/MDA5 DKO (red). d PFU per mg of tissue for spleens and brains from the mice assayed in previous panels. Each symbol represents a single organ from WT mice (blue) or RIG-I/MDA5 DKO mice (red). The lines are the means ± SEM. n = 3 mice per group per genotype. e WT (blue) or RIG-I/MDA5 DKO (red) BMMs were in vitro polarized and infected with WNV. FFU per mL of supernatant are shown. f RNA from in vitro polarized, WNV-infected WT (blue) or RIG-I/MDA5 DKO (red) BMMs was assayed for genomic copies of WNV. For panels e, f, bars are means ± SEM. For panels b, c, e, f, data are combined from n = 3 independent experiments, and lines are the means ± SEM with the following total mouse numbers: Mock WT n = 6; Mock DKO n = 2; WNV WT n = 11; WNV DKO n = 6. Stars indicate statistical significance between the marked groups (Welch’s unpaired t-test). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Source data are provided as a Source Data file