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. 2023 Nov 9;13:19440. doi: 10.1038/s41598-023-46254-1

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© The Author(s) 2023

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Inhibition of TLR1/2 signaling remarkably impacts the infiltration of immune cells after immunogenic chemotherapy treatment in vivo. (A) The effector/memory cells of CD3⁺CD45⁺ tumor-infiltrating immune cells were analyzed by flow cytometry (n = 4–5). The percentage of CD8⁺ T_EM cells (CD44⁺CD62L^-CD8⁺CD3⁺CD45⁺ T cells) in total T cells (CD3⁺CD45⁺ T cells) was examined. One-way ANOVA t test, *p < 0.05 and **p < 0.01. (B) Gating strategy of the IFNγ⁺CD8⁺ tumor-infiltrating immune cell profile. (C) IFNγ⁺CD8⁺ tumor-infiltrating immune cells were analyzed by flow cytometry (n = 4–5). The percentage of IFNγ⁺CD8⁺ T cells (IFNγ⁺CD8⁺CD3⁺CD45⁺ T cells) in total T cells (CD3⁺CD45⁺ T cells) was examined. One-way ANOVA t test, *p < 0.05 and **p < 0.01. (D) The Foxp3⁺ T regulatory cells were analyzed by flow cytometry (n = 4–5). The percentage of Foxp3⁺Treg cells (Foxp3⁺CD4⁺CD25⁺ T cells) in total T cells (CD3⁺CD45⁺ T cells) was examined. One-way ANOVA t test. (E) The density of tumor-infiltrating CD8 cells within the tumor microenvironment in patients with locally advanced colon carcinoma was analyzed by immunohistochemistry. The association between CD8 density and TLR1-N248 polymorphism was analyzed (n = 410, p = 0.0455). Unpaired t test.