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. 2022 Oct 24;25(11):1446–1457. doi: 10.1038/s41593-022-01183-6

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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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Fig. 2 — a, Immunofluorescence staining and quantification of C4b, Serpina3n, B2m and STAT1 in CC1⁺ oligodendrocytes in the white matter of 3- and 24-month-old mice (C4b⁺CC1⁺, 3-month, n = 3, 24-month, n = 5, ^*P = 0.0357; Serpina3n⁺CC1⁺, 3-month, n = 6, 24-month, n = 4, ^**P = 0.0095; B2m⁺CC1⁺, 3-month, n = 4, 24-month, n = 4, ^*P = 0.0286; STAT1⁺CC1⁺, 3-month, n = 5, 24-month, n = 5, ^**P = 0.0079; data are mean ± s.e.m.; P values are from a two-tailed Mann–Whitney U-test). Scale bar, 20 µm; for B2m, 10 µm. b, Immunofluorescence showing CD3⁺CD8⁺ T cells (indicated by arrowheads). Scale bar, 20 μm. Quantification of CD3⁺ T cells and CD8⁺ T cells in the gray (GM) and white (WM) matter of 3- and 24-month-old mice (n = 4 mice per group, 3-month WM versus 24-month WM, CD3⁺, ^***P = 0.0003, CD8⁺, ^***P = 0.0005; 24-month GM versus 24-month WM, CD3⁺, ^**P = 0.0050, CD8⁺, ^**P = 0.0032; data are mean ± s.e.m.; P values represent a two-sided Student’s t-test). c, Immunofluorescence of CD8⁺ T cells and STAT1⁺CC1⁺ oligodendrocytes in proximity in the white matter of 24-month-old mice. Scale bars, 20 μm. Bar plots show quantification of STAT1⁺ and STAT1⁻CC1⁺ proximity to CD8⁺ T cells and vice versa (3 sections per mouse were selected; a total of 134 CD8⁺ T cells and 272 STAT1⁺CC1⁺ oligodendrocytes from 4 mice were analyzed). d, Quantification of the percentage of STAT1⁺CC1⁺ oligodendrocytes found in proximity to random cells compared with CD8⁺ T cells (n = 4 mice per group, ^**P = 0.0052; data are mean ± s.e.m; P value represents a two-sided paried Student’s t-test). e, Immunofluorescence staining and quantification of CD8⁺ T cells, CD4⁺ T cells, B2m⁺ and STAT1⁺CC1⁺ oligodendrocytes in the white matter of mice treated with anti-PD-1 and CTLA-4 (ICB) and isotype control antibodies (CTR) for 6 weeks starting at an age of 18 months (CD8⁺, n = 4, ^**P = 0.0011; CD4⁺, n = 3; STAT1⁺CC1⁺, n = 4, ^*P = 0.0244; B2m⁺CC1⁺, n = 4, ^*P = 0.0286; data are mean ± s.e.m.; P values represent a two-sided Student’s t-test (CD8⁺, STAT1⁺CC1⁺) or two-tailed Mann–Whitney U-test (CD4⁺, B2m⁺CC1⁺)). Scale bar, 20 µm. NS, not significant.

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