Extended Data Figure 2. Analysis of immune cells in P6 and P11 Krit1^fl/fl and iECre;Krit1^fl/fl brains.

a, Gating strategy for B cells, NK cells, γδ T cells, CD4 T cells, CD8 T cells, eosinophils, neutrophils and monocytes/macrophages from cerebrum and cerebellum is shown. Cellular surface markers used were as follows: Neutrophils (CD45⁺, CD11b⁺, Ly6-G⁺), Eosinophils (CD45⁺, CD11b⁺, CD11c⁻, Ly6G⁻, Siglec-F⁺, SSChi), Monocyte/Macrophage (CD45⁺, CD11b⁺, CD11c⁻, Ly6G⁻, Siglec-F⁻, SSClo), NK cells (CD45⁺, CD11b⁻, CD19⁻, NK1.1⁺), B cells (CD45⁺, CD11b⁻, NK1.1⁻, CD19⁺), γδ T cell (CD45⁺, CD11b⁻, NK1.1⁻, CD19⁻, CD3⁺, TCRγδ⁺), CD4 T cell (CD45⁺, CD11b⁻, NK1.1⁻, CD19⁻, CD3⁺, TCRγδ⁻, CD8⁻, CD4⁺), CD8 T cell (CD45⁺, CD11b⁻, NK1.1⁻, CD19⁻, CD3⁺, TCRγδ⁻, CD4⁻, CD8⁺). b, The number of B cells, NK cells, γδ T cells, CD4 T cells, CD8 T cells, eosinophils, neutrophils, and monocytes/macrophages isolated from P6 cerebrum (top) and cerebellum (bottom) is shown for susceptible Krit1^fl/fl and iECre;Krit1^fl/fl littermates. N≥6 per group. No significant differences were detected. c, The number of B cells, NK cells, γδ T cells, CD4 T cells, CD8 T cells, eosinophils, neutrophils, and monocytes/macrophages isolated from P11 cerebrum (top) and cerebellum (bottom) is shown for susceptible Krit1^fl/fl and iECre;Krit1^fl/fl littermates. N≥6 per group. d–e, Frequency of RORγt⁺ CD4 T cells isolated from P6 and P11 cerebellum. N≥6 per group. Error bars of all graphs shown as s.e.m. and significance determined by unpaired, two-tailed Student’s t-test. *indicates p<0.05. Note that there is significant immune cell presence in the cerebellum of susceptible iECre;Krit1^fl/fl animals at P11 but not at P6.