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. 2020 Jul 17;11:3584. doi: 10.1038/s41467-020-17414-y

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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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Fig. 3 — a Line plot for flow cytometry quantification of PD-1⁺CTLA-4⁺ live CD8⁺ T-cells in tumors pre and post therapy on cycle 3. Responders are indicated with an empty square, while non-responders with a full circle (n = 15). Flow cytometry quantification of CD3⁺ (b) and CD45⁺ cells (c) at baseline and post treatment. d Flow cytometry quantification of CD4⁺ and CD8⁺ in the tumor microenvironment in non-responders (NR) versus responders (R). Flow cytometric quantification of HLA-DR (e) and Ki67 (f) expression in live CD8⁺ T-cells from metastatic tumors before (Baseline) and after 3 cycles of treatment (Cycle 3) (n = 14). The markers represent the minimum, first quartile, median, third quartile, and maximum values, respectively (e, f). Data are presented as the mean ± SD. Statistical significance is indicated by p-values as *p ≤ 0.05; **p ≤ 0.01; ****p ≤ 0.0001; NS = not significant. This was assessed by two-tailed Wilcoxon text for a–c and e, f and by unpaired T test for D.