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. 2022 Sep 24;13:5606. doi: 10.1038/s41467-022-33067-5

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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. 5 — a Gene expression of interferon gamma receptor 1 (IFNGR1) in isolated human regulatory T cells (Tregs) from visceral adipose tissue (VAT) and peripheral blood (PB) of lean versus obese subjects (VAT: N = 5 lean, N = 7 obese; PB: N = 6 lean, N = 7 obese). b Gene expression of IFNGR1 in isolated human Tregs from VAT vs. PB. c Gene expression of programmed cell death protein 1 (PDCD1 or PD-1), T cell immunoreceptor with Ig and ITIM domains (TIGIT), OX40, LKB1, Annexin 5 (ANXA5), and Caspase (CASP) 8 and 9 in isolated human PB Tregs (N = 4 per group) cultured for 24 hr and 72 hr, with or without recombinant IFNγ (20 ng/ml); **P < 0.010; *P < 0.05. d Gene expression in human cultured subcutaneous adipocytes of the PD-1 ligands PD1GL1 (CD274) and PDCD1LG2. Adipocytes were pretreated with a JAK inhibitor (JAK I) for 1.5 hr in adipocyte maintenance medium prior to co-treatment with IFNγ (20 ng/ml) for 48 hr (n = 3). Fold changes vs. vehicle samples are not shown in the graph and all close to 1; ***P < 0.001. e VAT Tregs and PB Tregs in lean (N = 6) and obese (N = 12) human subjects positive for PD-1, TIGIT, and CTLA4 and in lean (N = 1) and obese (N = 7) human subjects positive for OX-40 by flow cytometry; ****P < 0.0001; ***P < 0.001; **P < 0.010. f Gene expression of LKB1 in human VAT (N = 13) vs. PB (N = 13) from the same individuals; **P < 0.010. g % proliferation in cultured activated peripheral blood mononuclear cells (PBMCs) without Tregs (first panel) and with flow sorted Tregs from PB (second panel) and VAT (third panel) in a representative obese patient. h % inhibition of proliferation in cultured activated peripheral blood mononuclear cells (PBMCs) with Tregs flow sorted from PB (N = 14) vs. VAT (N = 14) from the same individuals, ***P < 0.001. i Correlation between % of total Tregs positive for PD-1 vs % inhibition of proliferation in cultured PBMCs with flow sorted Tregs from VAT and PB. Source data are provided as a Source Data file. Group comparison data was analyzed by two-sided, independent T tests without correction for multiple variables and presented as mean ± SEM.