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. 2022 May 26;23(6):878–891. doi: 10.1038/s41590-022-01208-z

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Extended Data Fig. 2 — a, Spleen from wildtype and Foxp3IL-2 mice were compared by flow cytometry (n = 4, 6). Frequency of key markers (CD25, CD44, CD62L, CD103, CTLA4, Helios, ICOS, Ki67, KLRG1, Neuropilin1, PD1, ST2, Tbet) on Treg cells in spleen. b, Brain from wildtype and αCamKII^IL-2 mice were compared by high-dimensional flow cytometry (n = 4, 3). Frequency quantification of key markers on Treg cells. c, tSNE of blood, spleen and brain Treg cells built on key markers. Colors indicate annotated FlowSOM clusters, with quantification in Supplementary Fig. 5 f. tSNE run on samples pooled post-acquisition, with quantification performed on individual samples (n = 4, 3). The residential cluster is characterized as CD25hiCD69 + PD1 + CD103 + . d, tSNE of total leukocytes from brain of wildtype and αCamKII^IL-2 mice, built on lineage markers with quantification. tSNE run on samples pooled post-acquisition, with quantification performed on individual samples (n = 4, 3). e, NK, CD4 and CD8 T cells, (left) as a proportion of CD45 + CD11b⁻ cells in the brain (n = 4, 3)., and (right) in absolute numbers, together with Treg cells (n = 6, 4). f, tSNE of brain CD4 conventional T cells built on key markers (CD62L, CD44, CD103, CD69, CD25, PD-1, Nrp1, ICOS, KLRG1, ST2, Ki67, Helios, T-bet, CTLA4). tSNE run on samples pooled post-acquisition, with quantification performed on individual samples (n = 4, 3). Colors indicate annotated FlowSOM clusters, with quantification and g, frequency of marker expression. h, tSNE of brain CD8 T cells built on key markers. tSNE run on samples pooled post-acquisition, with quantification performed on individual samples (n = 4, 3). Colors indicate annotated FlowSOM clusters, with quantification and i, frequency of marker expression. j, tSNE of brain NK cells built on key markers (t-SNE run on samples pooled post-acquisition, with quantification performed on individual samples (n = 4, 3). Colors indicate annotated FlowSOM clusters, with quantification and k, frequency of marker expression. Data are displayed as mean ± s.e.m. (a,b,d-k). Statistical analyses were performed using multiple unpaired two-tailed Student’s t-test.

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