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. 2020 Apr 10;10:6220. doi: 10.1038/s41598-020-63060-1

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© The Author(s) 2020

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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Co-expression pattern of PD-1 and TIM-3 on both CD8 and CD4 T cells of peripheral blood mononuclear cells (PBMC), adjacent normal tissue-infiltrating lymphocytes (NILs), and tumour tissue-infiltrating lymphocytes (TILs) in 48 patients with renal cell carcinoma. (a) Gating strategy to identify subsets of CD8 and CD4 T cells within PBMC, NILs, and TILs. CD8 T cells were classified into four groups (Fr. I: PD-1-TIM-3−, Fr. II: PD-1lowTIM-3−, Fr. III: PD-1+TIM-3−, Fr. IV: PD-1+TIM-3+), and CD4 T cells were classified into 5 groups (Fr. I: PD-1-TIM-3−, Fr. II: PD-1lowTIM-3−, Fr. III: PD-1+TIM-3−, Fr. IV: PD-1+TIM-3+, Fr. V: PD-1lowTIM-3+). (b) Expression ratios of each fraction on both CD4 and CD8 T cells were compared among NILs and TILs simultaneously extracted from the same patients. Comparison of two sample types was performed by Mann-Whitney U test. The central tendency of the box plot shows the median of each group, and the upper and lower ranges of the box plot show the 25^th and 75^th percentiles of each data set, respectively. **P < 0.01, *P < 0.05.