Fig. 5

The identified microglial subsets are not peripherally derived nor represent region-specific phenotypes. a-d Microglia ‘subsets’ gated and overlaid onto tSNE plots representing WNV dpi 7 brains from animals treated with isotype (a) and VLA4/CCL2/Ly6C blockade antibodies (b) and mock-infected brains from animals treated with (d) and without (c) PLX5622. e, f IMC images of the hippocampus (e) and mid-brain (f) from a WNV dpi 5 animal, showing the expression of CD45 (cyan), Iba-1 (green), CD86 (yellow) and Ly6C (red). Circles show the different microglia ‘subsets’ P2RY12^hi CD86⁺ (dark purple), P2RY12^lo CD86⁻ (light green), P2RY12^lo CD86⁺ (light purple), P2RY12^hi CD86⁻ (dark green). g tSNE plots clustered on live myeloid cells (UVLD⁻ CD11b⁺ CD45⁺) from WNV dpi 7 brains, segmented into 5 anatomical regions. Myeloid populations from the olfactory bulb, frontal cortex, posterior cortex, pons/medulla and cerebellum were gated and overlaid onto the plot. h, i FACs plots and pie graphs showing the relative proportions of each microglia ‘subset’ found in 5 anatomical regions of non-infected (h) and WNV-infected brains (i). Values on pie graphs indicate the frequency of each microglial population out of total microglia. Data shown in a-d is representative of two independent experiments with six mice in each group. Data shown in g-i is representative of one experiment with six brains pooled per brain region