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. 2019 Sep 11;7:246. doi: 10.1186/s40425-019-0725-7

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© The Author(s). 2019

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Fig. 4 — | Combining CAI with DMF or 1-MT increased the number of tumor-infiltrating CD8⁺ T cells and downregulated PD-1 expression. BALB/c or C57BL/6 mice were subcutaneously injected with 1 × 10⁶ tumor cells (C26 cells, B16 cells or 4 T1 cells, n = 6 in every group) and received the indicated drugs for 7 days after the day the tumor size reached 5 mm in diameter. T cells in the tumor microenvironment were sorted and analyzed by flow cytometry. a Representative flow cytometry plots showing the fraction of CD8⁺ T cells within the CD3⁺ TILs in the C26, B16- or 4 T1- tumor microenvironment. b CD8⁺ T cell numbers per gram of tumor in different groups. c The percentage of PD-1⁺CD8⁺ T cells within TILs in the tumor microenvironment. d, e and f The same measurements from the evaluation of the effects of another drug combination (CAI and 1-MT) in C26-, B16- or 4 T1- tumor bearing mice. Data are from three independent experiments, and the error bars represent the mean ± s.e.m. **p < 0.01 by one-way ANOVA (a-f)