Extended Data Fig. 6: Generation of Ifnγr receptor deficient SJL mice.

(a) Schematic of CRISPR/Cas9⁻based Ifnγr1 targeting strategy to delete 203 bp spanning the promoter and first exon of the Ifnγr1 gene. (b) Schematic showing the WT and mutant Ifnγr1 locus with details of the respective 203 bp deletion. (c) Genomic PCR analysis of Ifnγr1 ^−/− SJL mouse (~400 bp) and WT control (~600 bp). Representative out of infinite number of experiments. (d) Flow cytometric analysis of non-classical CD115⁺ Ly6c- blood monocytes for expression of CD119 (Ifnγr) in WT, Ifnγr1^−/+ or Ifnγr1 ^−/− SJL mouse. e, f. Flow cytometric analysis of IFNγR (E) or MHC-II (F) expression of MoMF (gated as live CD45^hi CD11b⁺ TMEM119^-) in control or Mg^ΔIfnγr1 mice at REM. MFI, mean fluorescent intensity. N = 5 individual mice per group, lines represent mean. Significance was calculated using two tailed t-test. Dashed line/empty circles - FMO; black histogram/black dots - control; red histogram/red triangles - Mg^ΔIfnγr1 mice. g. Normalized MFI of Treg cells markers on Foxp3⁺ T cells of Mg^ΔIfnγr1 and control brains. Data combined from 5 independent experiments n = 27 individual mice per group, lines represent mean. Significance was calculated using two tailed t-test. *** p < 0.001. h. Overlay of microglia (light blue; gated on live Ly6c/g1⁻ CD11b⁺ CD45^int TMEM119⁺), singlet T cells (red, gated on live Ly6c/g1^- CD11b⁻ CD45^hi CD4⁺ TCRβ⁺) and total T cells (gray; gated on live Ly6c/g1⁻ CD45^hi CD4⁺ TCRβ⁺), related to Fig. 5g. i. Normalized MFI of non-Treg cells (CD4⁺TCRβ⁺ Foxp3⁻) of Mg^ΔIfnγr1 and control brains. Data combined from 5 independent experiments n = 27 per group, lines represent mean. Significance was calculated using two tailed t-test. *p < 0.05, *** p < 0.001.