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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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Extended Data Fig. 3 — a, Immunofluorescence staining showing the different densities of CD8⁺ T cells, STAT1⁺CC1⁺ cells, MHC1⁺IBA1⁺ cells and nodules (Galectin-3⁺Iba1⁺ cell clusters) between frontal white matter away from lateral ventricles and medial white matter close to lateral ventricles. Scale bar 20 µm. b, Quantification of CD8⁺ T cells, STAT1⁺CC1⁺ cells, MHC1⁺IBA1⁺ cells, and nodules (Galectin-3⁺IBA1⁺ cells cluster) density difference between frontal white matter and medial white matter of 24-month old mice (for CD8⁺, STAT1⁺CC1⁺, and nodules, n = 4 mice per group; CD8⁺, *P = 0.0438; STAT1⁺CC1⁺, *P = 0.0148; nodules, **P = 0.0088; for MHC1⁺IBA1⁺, n = 3 mice per group; *P = 0.0479; data are means±s.e.m. P values represent a two-sided paried Student’s t-test). c, T cells proportion per (n = 14 independent experiments) in the Tabula Muris Senis dataset. The central line denotes the median, boxes represent the interquartile range (IQR), and whiskers show the distribution except for outliers. Outliers are all points outside 1.5 times of the IQR. d, Representative confocal images of aged white matter stained for CD3 (red) and CD8 (green). The experiment was repeated three times independently with similar results. Scale bar, 20 µm. e, Immunofluorescence image showing CD8⁺ T cells (white) and Serpina3n⁺ (red) in CC1⁺ oligodendrocytes (green) in the white matter of 24-month old mice. Scale bars, 20 μm. Quantification of the percentage of Serpina3n⁺CC1⁺ oligodendrocytes found around random cells (<20 µm) compared to the percentage of Serpina3n⁺CC1⁺ oligodendrocytes found around CD8⁺ T cells (n = 3 mice per group; data are means±s.e.m.; P value represents a two-sided paried Student’s t-test). f, Immunofluorescence showing expression of Serpina3n (green) in CC1⁺ oligodendrocytes (red) in the white matter of mice treated with antibodies anti PD-1 and CTLA-4 and isotype control for 6 weeks starting at an age of 18 months. Scale bar 20 µm. Quantification of CC1⁺ oligodendrocytes expressing Serpina3n in the white matter of mice treated with antibodies anti PD-1 and CTLA-4 and isotype control antibodies (n = 4 mice per group; data are means±s.e.m).

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