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. 2022 Jul 1;13:3799. doi: 10.1038/s41467-022-31135-4

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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. 6 — Ldlr^−/− and T-Abc^dkoLdlr^−/− mice (a–e, i–j) and control (Ldlr^+/+) and T-Abc^dko mice (f–h) were fed a chow diet for 12–13 months, and wild-type mice for 3 months (young) or 24 months (aged) (i–j). a–j Spleens were collected and CD4⁺ and CD8⁺ T cells were isolated. a, b CD4⁺ and CD8⁺ T cells were stimulated with αCD3/IL-2 for 12 h with concomitant staining for cleaved caspase3/7. a Representative flow cytometry plots of cleaved caspase 3/7 in CD4⁺ (p = 0.015) and CD8⁺ (p = 0.007) T cells gated as in Supplementary Fig. 12d and (b) quantification. n = 6 Ldlr^−/− and n = 5 T-Abc^dkoLdlr^−/− mice. c–h T cells were labeled with CFSE and stimulated with αCD3/αCD28 beads. CFSE dilution was measured by flow cytometry at 72 h after stimulation. c, f Representative CFSE dilutions gated as in Supplementary Fig. 11c. The number of divisions were quantified for (d) CD4⁺ (division 0, p = 0.000017; 2, p = 0.00002; 3, p < 0.000001; 4, p < 0.000001) and (e) CD8⁺ (division 0, p = 0.0029; 1, p = 0.00095; 3, p = 0.0023; 4, p = 0.0000097; 5, p = 0.008) T cells. n = 7 Ldlr^−/− and n = 6 T-Abc^dkoLdlr^−/− mice. The number of divisions were quantified for (g) CD4⁺ (division 0), p = 0.0002; 1, p = 0.013; 2, p = 0.017; 3, p = 0.00067; 4, p = 0.0031 and (h) CD8⁺ T cells (division 0, p = 0.0004; 1, p = 0.0056; 3, p = 0.012; 4, p = 0.0003; 5, p = 0.02). n = 4 control and n = 4 T-Abc^dko mice. i–j T cells were isolated and RNA was extracted. Cyclin dependent kinase inhibitor 1a (Cdkn1a) mRNA expression was measured in (i) CD4⁺ (Ldlr^−/− or aged wild-type vs T-Abc^dkoLdlr^−/−, p < 0.0001) and (j) CD8⁺ (Ldlr^−/− vs T-Abc^dkoLdlr^−/−, p=0.0003; aged wild-type vs T-Abc^dkoLdlr^−/−, p < 0.0001) T cells by qPCR and shown as fold change compared to young wild-type mice. n = 8 young wild-type, n = 6 aged wild-type, n = 6 Ldlr^−/−, and n = 5 T-Abc^dkoLdlr^−/− mice. For all panels, error bars represent SEM. Biologically independent samples were included. p value was determined by unpaired two-tailed Student’s t test (b, d–e, g, h) or one-way ANOVA with Bonferroni post-test (i, j). *p < 0.05, **p < 0.01, ***p < 0.001. Source data are provided as a Source Data file.