Extended Data Fig. 2. Meningeal γδ T cells expression profile and additional behavior paradigms using γδ T cell-deficient mice.

a, Normalized expression of Tcrgv6 transcripts (Vγ4 chain, Garman nomenclature) in meningeal and splenic γδ T cells obtained by single cell RNA-Seq. b, Graph bars of the TCR Vγ-usage in γδ T cells. b_i, Meningeal and b_ii, splenic γδ T cells. b_iii, Overview of Vγ-chain usage (Vγ1, Vγ2, Vγ3 or others (Vγ4 or Vγ5)). c_i, Representative contour plots of the intracellular staining for IL-17a and IFNγ in αβ T cells (TCR β⁺NK1.1^neg) and γδ T lymphocytes (Vγ1, Vγ2, Vγ3–negative or positive fractions) isolated from steady state meninges. c_ii-v, Graph bars of the percentage of IL-17a– or IFNγ– producing cells in these different meningeal T cell subsets. One-way ANOVA followed by Bonferroni's multiple comparisons test. d, UMAP projection of γδ T cells colored by the scaled expression of Slc6a13 (GAT2). e, Average expression of chemokines in CD45⁺ cells isolated from steady state dura mater obtained from public database¹. f, Representative histogram of the expression of CXCR6-GFP in meningeal T cell subsets. g, Absolute numbers of conventional αβ T cells isolated from CXCR6–sufficient and –deficient mice. Unpaired two-tailed t test. Data represent one independent experiment. h_i, Adult Ccr2^CreERT2:Ai6 mice were injected daily with tamoxifen (i.p., 10 mg/kg) and three days later meninges, spleen and blood were harvested for FACS analysis. Representative histograms of CCR2-ZsGreen expression in γδ T cells and conventional αβ T lymphocytes by flow cytometry. h_ii, Graph bars of the percentage of γδ T cell– and h_iii, αβ T cell– expressing CCR2-ZsGreen in spleen, blood and meninges. h_iv, Histograms of the expression of CCR2-ZsGreen in meningeal γδ and αβ T lymphocytes. h_v, Frequency of CCR2–expressing cells. One-way ANOVA followed by Bonferroni's multiple comparisons test. i_i, Representative dot plots of the percentage of meningeal TCR γδ– and TCR β– expressing cells isolated from WT or Ccr2^−/− (Ccr2^{RFP/ RFP}) mice. i_ii, Percentage and i_iii, absolute number of meningeal γδ T cells. i_iv, Percentage and i_v absolute number of meningeal αβ T cells. Unpaired two-tailed t test. j_i, Dot plots of the frequency of inflammatory monocytes (CD11b^hiLy6C^hi) isolated from WT or CCR2-deficient mice. j_ii, Percentage and j_iii, absolute number of meningeal inflammatory monocytes. Data are pooled from two independent experiments with similar results. Unpaired two-tailed t test. k, Social interaction of wild-type and Tcrd^−/− mice was assessed in the three-chamber test and the percentage of time investigating either a mouse (social) or an object (inanimate) was calculated. WT (n = 9); Tcrd^−/− (n = 11). Two-way ANOVA followed by Bonferroni's multiple comparisons test. l_i, For the spatial memory task, WT and γδ T cell-deficient mice were assessed in the training and l_ii, novel location tasks of the novel location recognition (NLR) test and the time spent with either a familiar or novel object was determined. WT (n = 8); Tcrd^−/− (n = 8). Two-way ANOVA followed by Bonferroni's multiple comparisons test. m, Foraging behavior of wild-type and Tcrd^−/− mice assessed in the exploratory and n, foraging phase of the foraging paradigm. Data is pooled from three independent cohorts. WT (n = 18); Tcrd^−/− (n = 18); Unpaired two-tailed t test. Data are shown as mean ± s.e.m. and each dot symbol represents individual mouse.