Extended Data Fig. 8: Depletion strategy of ME-Macs.

a, Schematic of α-CSF1R (AFS98) and α-CCR2 (MC21) treatment. Tissues were analysed 24 h and 5 d post-treatment, at site of injection (local) or 3 cm distant (distal). b, FACS gating strategy of immune cell phenotyping (macrophages, monocytes, B cells, eosinophils, neutrophils and DC) in ME, LP, blood and dura mater post- α-CSF1R/ α-CCR2 treatment vs IgG. c, Absolute cell counts of CX3CR1⁺CD11b⁺Ly6C^loMHCII^hi ME-Macs, CX3CR1⁺CD11b⁺Ly6C^hiMHCII^lo monocytes, CD19⁺ B cells, CD11b⁺Ly6G⁺ neutrophils, CD11b⁺SiglecF⁺ eosinophils in ME and LP; CD11c⁺CD103⁺ DC in LP; CX3CR1⁺CD11b⁺Ly6C^hi monocytes^, CD3⁺ T cells, CD19⁺ B cells, CD11b⁺Ly6G⁺ neutrophils and CD11b⁺SiglecF⁺ eosinophils in blood; CX3CR1⁺CD11b⁺Ly6C⁻ macrophages, CD11b⁺Ly6C⁺ monocytes and CD11c⁺CD103⁺ DC in dura mater. n = 4 mice for IgG (except n = 3 for ME macrophages, local and distal 5 d; ME monocytes, local and distal 5 d; LP B cells and eosinophils local 5 d; ME B cells, local and distal 5 d; ME neutrophils, local and distal 5 d; LP B cells and eosinophils, distal 5 d, ME eosinophils, local and distal 5 d, LP macrophages local and distal 5 d; LP neutrophils, monocytes and dendritic cells, local 5 d; LP monocytes, dendritic cells and neutrophils, distal 5 d; blood monocytes, T cells, B cells eosinophils, neutrophils, 5 d; dura macrophages, monocytes and dendritic cells 5 d. n = 3 mice for α-CSF1R, α-CCR2 and α-CSF1R/α-CCR2 per time point. 1 experiment per time point, data analysed using 1way ANOVA with Bonferroni’s multiple comparison test if applicable, Kruskal-Wallis with Dunn’s multiple comparison test or Welch’s 1way ANOVA with Dunnett’s multiple comparison test. d, FACS plots of CD11b⁺CX3CR1⁺ microglia and CD163⁺ BAM at 5 d post treatment. Data are mean ± s.e.m.

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