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. 2021 Sep 6;12:5303. doi: 10.1038/s41467-021-25645-w

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© The Author(s) 2021, corrected publication 2024

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 Female C57BL/6 mice were cohoused with outbred, wild-type pet shop mice in large rat cages, separated by a perforated barrier. After >60 days of cohousing expression of Cd49d, CD69, KLRG1, CD27, and CD25 was analyzed by flow cytometry on subpopulations of Tn CD8⁺ T cells from lymph nodes of CH mice and SPF controls (Color scheme: red-Tn Ly6C⁻ from SPF mice; blue-Tn Ly6C⁺ from SPF mice; orange-Tn Ly6C⁻ from CH mice; light green-Tn Ly6C⁺ from CH mice; dark green-CD44^hi from SPF mice; brown-CD44^hi from CH mice). B Upregulation of Ly6C was upregulated on central memory (CM) cytotoxic T cells (CD8⁺CD44^hiCD62L^hi), CD44^negCD62L^hi and CD44^intCD62L^hi cells (n = 4 SPF mice, n = 5 CH mice per group, data presented as mean ± sd, ***p < 0.001). C Ly6C was upregulated on Tn CD8⁺ cells in the blood, lymph nodes, and spleen (n = 5 mice per group, data presented as mean ± sd, ****p < 0.0001). D Tn CD8⁺ cells were magnetically enriched^, CFSE labeled and transferred into CH mice for 14 days (n = 7 ctrl mice, n = 8 CH mice, data presented as mean ± sd, ***p = 0.0003). E Expression of CD5 and SCA-1 on Ly6C ± CD8⁺ Tn from SPF and CH (Color scheme^: red-Tn Ly6C⁻ from SPF mice; blue-Tn Ly6C⁺ from SPF mice; orange-Tn Ly6C⁻ from CH mice; light green-Tn Ly6C⁺ from CH mice). F Ly6C ± CD8⁺ Tn from SPF and CH (n = 5 mice per group, data presented as mean ± sd, left to right *p = 0.0481, *p = 0.0336) were stained intracellularly for expression of Eomes and (G) BCL-2 (n = 5 mice per group, data presented as mean ± sd, *p = 0.0168). A, B, C, E, F, and G data are representative of two experiments (with n = 5 mice per group). D Data are pooled from two experiments (total N = 7 mice per group (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001). D Two-tailed Mann-Whitney U-test; A, B, C, E, F, and G) one-way ANOVA, with Sidak post hoc correction.