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. 2019 May 30;9:8065. doi: 10.1038/s41598-019-44523-6

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© The Author(s) 2019

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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Overexpression of cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) decreases the frequency of CD4⁺ T cells and CD4⁺ forkhead box P3 (Foxp3)⁺ regulatory T cells (Tregs) in angiotensin II-treated or saline-treated mice. Seven days after the pump implantation, lymphoid cells from spleen and lymph nodes (LNs) were prepared. Apolipoprotein E-deficient (Apoe^−/−) mice infused with angiotensin II or saline served as controls. (A) Representative results of intracellular CTLA-4 expression in splenic CD4⁺Foxp3⁺ Tregs and CD4⁺Foxp3⁻ T cells of angiotensin II-infused Apoe^−/− and CTLA-4-Tg/Apoe^−/− mice assessed by flow cytometry. The expression levels of intracellular CTLA-4 were examined. The data are shown as mean fluorescence intensity (MFI). n = 5 per group. (B) Representative results of Foxp3 and CD4 expression in the spleen of angiotensin II-infused Apoe^−/− and CTLA-4-Tg/Apoe^−/− mice assessed by flow cytometry. The graphs represent the percentage of CD4⁺ T cells or CD4⁺Foxp3⁺ Tregs within spleen and LN cells or spleen and LN CD4⁺ population, respectively. n = 5 mice per group. Data points represent individual animals. Horizontal bars represent means. Error bars indicate s.d. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; one-way ANOVA followed by Tukey’s post hoc test. Ang-II, angiotensin II; SSC, side scatter.