Fig. 9

Microglia are the principal producers of IL-12 late in WNV-infection. a Heatmap showing the proportion of cells that are IFN-γ⁺, IL-12/IL23 (p40)⁺, DAF-FM⁺ or CD206⁺ in WNV-infected dpi 7 brains. b, c tSNE plots showing the expression of P2RY12 (b) and IL-12/IL-23 (p40) (c) on live cells isolated from WNV-infected brains at dpi 7. Red circles indicate where microglia are positioned on plots. d RNAse protection assays (RPA) performed for IL-12 (p35), IL-23 (p19), IL-12/IL-23 (p40), TNF, IL-6, IFN-γ and IL-17A on mock and WNV-infected brains at dpi 7. Lanes 1-3 indicate individual animals in each group. RPL32-4A was used as an internal control for RNA loading during RPA analysis. e, f The number (e) and proportion (f) of total microglia expressing IL-12/IL-23 (p40) at dpi 0, 5 and 7. g, h Proportion of P2RY12^hi CD86⁺ (dark purple), P2RY12^lo CD86⁻ (light green), P2RY12^lo CD86⁺ (light purple) and P2RY12^hi CD86⁻ (dark green) expressing IL-12/IL-23 (p40) at WNV dpi 7 out of the total microglia population (g) and within microglia sub-populations (h). *P < 0.0332, **P < 0.0021, Kruskal-Wallis test and Dunn’s multiple comparisons test