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. 2021 Jul 22;12:4474. doi: 10.1038/s41467-021-24734-0

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© The Author(s) 2021

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. 4 — a Representative CD44/CD62L staining of CD45⁺TCRβ⁺NK1.1⁻CD8α⁺ gated liver MNCs (upper) and KLRG1/CD69 staining of CD45⁺TCRβ⁺NK1.1⁻CD8α⁺CD44⁺CD62L⁻ gated (Tem + Trm) liver MNCs (lower) of RES (5 weeks) mice. b Frequency of each CD8⁺ T cell subset in CD45⁺ liver MNCs derived from the indicated groups of mice (n = 6 mice for ND and HFHC groups, and n = 8 mice for RES group). Data are presented as mean ± SEM. One-way ANOVA with Tukey’s multiple comparisons post-hoc test was applied. c Relative abundance of partner or parent cells in the blood and liver MNCs of mice two weeks following the parabiosis surgery between Ly5.2⁺ RES (5 weeks) mice and Ly5.1⁺ RES (5 weeks) mice (n = 6 pairs). Data are presented as mean ± SEM. d–f Bulk RNA-seq analysis of isolated liver naïve CD8⁺ T cells from ND mice (ND naïve; n = 3), CD8⁺ Tem cells from HFHC mice (HFHC Tem; n = 3), CD8⁺ Trm cells from HFHC mice (HFHC Trm; n = 3), and CD8⁺ Trm cells from RES (5 weeks) mice (RES Trm; n = 3). d Heat map of key upregulated (upper) or down-regulated (lower) genes characteristic of Trm. e K-means clustering analysis. Most variable 1200 genes were clustered to six groups. Featured genes determined by scRNA-seq were shown. f PCA analysis plot of top 1000 genes. g–i ScTCR-seq analysis of combined liver or spleen CD8 T cells derived from ND and RES (5 weeks) mice. g UMAP projection of combined liver and spleen CD8⁺ T cells (n = 53,659) derived from ND(Liver), ND (Spleen), RES (Liver), and RES (Spleen) mice. Green circle highlights clusters of Trm cells. h UMAP projection and i percentage of top 10 and other TCR clonotypes in each cell subset. Source data are provided as a Source data file.