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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. 4 — Splenic CD4⁺ a–f, CD8⁺ (a–f, j), or total T cells (g–i) were isolated from Ldlr^−/− and T-Abc^dkoLdlr^−/− mice. a–f CD4⁺ and CD8⁺ T cells were stimulated with αCD3/IL-2 for 12 h, fixed, permeabilized, stained for granzyme B (a, b), or interferon gamma (IFN-γ) (c, d), or alternatively, stained for lysosomal-associated membrane protein-1 (LAMP-1) (e, f). Gating strategies are shown in Supplementary Fig. 14. Representative flow cytometry plots of (a) CD8⁺granzyme B⁺ (p = 0.0096), (c) CD4⁺IFN-γ⁺ (p = 0.048) and CD8⁺IFN-γ⁺ (p = 0.002) T cells and (b, d) quantification as a percentage of CD8⁺ T cells after correction for isotype control (b) and as a percentage of CD4⁺ or CD8⁺ T cells after correction for fluorescence minus one (FMO) control (d). e Representative flow cytometry plots and (f) quantification of LAMP-1 expression on CD4⁺ and CD8⁺ (p = 0.027) T cells. b, f n = 7 Ldlr^−/− and n = 6 T-Abc^dkoLdlr^−/− mice. d n = 8 Ldlr^−/− and n = 5 T-Abc^dkoLdlr^−/− mice. g T cells were stimulated with αCD3/IL-2 for 12 h. Medium was collected and IFN-γ levels (p = 0.034) were measured by ELISA. n = 7 Ldlr^−/− and n = 6 T-Abc^dkoLdlr^−/− mice. h Wild-type bone marrow derived macrophages (BMDMs) were incubated with conditioned medium from CD8⁺ T cells stimulated with αCD3/IL-2 for 12 h, or recombinant IFN-γ (rIFN-γ; positive control) for 3 days. After medium removal, BMDMs were co-incubated with E. coli for 1 h, washed, incubated with gentamicin for 3 h, washed, and lysed. BMDM lysates were plated on agar overnight and E. coli colony forming units (CFU) were counted (Ldlr^−/− vs T-Abc^dkoLdlr^−/−, p = 0.0032; Ldlr^−/− or T-Abc^dkoLdlr^−/− vs rIFN-γ, p < 0.0001). n = 3 Ldlr^−/− and n = 3 T-Abc^dkoLdlr^−/− mice. i BMDMs were co-incubated with T cells in the presence of αCD3/IL-2 (to stimulate T cells) for 24 h. Medium was collected and endogenous lactate dehydrogenase (LDH) was assessed (p = 0.0023). 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 (a–i) or one-way ANOVA with Bonferroni post-test (j). *p < 0.05, **p < 0.01, ***p < 0.001. Source data are provided as a Source Data file.