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Fig. 8 | Journal of Neuroinflammation

Fig. 8

From: High-parameter cytometry unmasks microglial cell spatio-temporal response kinetics in severe neuroinflammatory disease

Fig. 8

Kinetics of microglia proliferation and death in the progression of lethal encephalitis. a, b Number (a) and percent (b) of total microglia in murine brains at dpi 0, 3, 4, 5, 6 and 7. *P < 0.0332, **P < 0.0021, ***P < 0.0002, ****P < 0.0001, Kruskal-Wallis test and Dunn’s multiple comparisons test. c, d Number (c) and proportion (d) of P2RY12hi CD86+ (dark purple), P2RY12lo CD86 (light green), P2RY12lo CD86+ (light purple) and P2RY12hi CD86 (dark green) microglia at WNV dpi 0, 4, 5, 6 and 7. *P < 0.0332, **P < 0.0021, ***P < 0.0002, ****P < 0.0001, two-way ANOVA with a Tukey’s multiple comparisons test. Data is presented as mean ± SEM and representative of 2-3 independent experiments. e, f Number (e) and percent (f) of P2RY12hi CD86+ (dark purple), P2RY12lo CD86 (light green), P2RY12lo CD86+ (light purple) and P2RY12hi CD86 (dark green) microglia that are BrdU+ in WNV-infected brains. *P < 0.0332, **P < 0.0021, ***P < 0.0002, two-way ANOVA with a Tukey’s multiple comparisons test. g FACs plots showing the expression of P2RY12 and BrdU on/in microglia in murine brains in WNE. h tSNE plots clustered on live cells from WNV-infected brains and overlaid with BrdU+ (proliferating) cells (red), which were determined with a fluorescence minus one (FMO) sample (i.e., no anti-BrdU staining). Microglia are outlined in blue on each plot. i FACs plots showing microglial cell expression of UVLD and Annexin V in WNV-infected brains. Values on FACs plots indicate the proportion of UVLD Annexin V, UVLD Annexin V+ and UVLD+ Annexin V+ microglia out of the total microglia population. j Number and percent of Annexin V+ and dead microglia in WNV-infected brains. k Heatmap showing the percent of UVLD+ and Annexin V+ myeloid and lymphoid populations in murine brains over the course of WNV. Data is presented as mean ± SEM and representative of at 1-2 independent experiments, with at least 4 mice per group

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