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. 2022 Apr 21;13(4):392. doi: 10.1038/s41419-022-04838-1

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© The Author(s) 2022

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. 5 — A The expression of MADCAM1 show a significant positive correlation with the fraction of lymphocyte and a negative correlation with the fraction of macrophages, in both TCGA cohort and FUSCC cohort. B Immunohistochemistry (IHC) analysis showed that the pericellular staining intensity of MADCAM1 in the tumor cells of samples carrying mutated MADCAM1 was significantly stronger than those carrying wild-type MADCAM1. Additionally, CD163 was significantly positively stained in the infiltrating immune cells of samples with mutated MADCAM1, when compared with those with wild-type MADCAM1. C Effect of MADCAM1^P270Q and MADCAM^D242N on stability of the Madcam1 protein. GC cells transfected vectors expressing wild-type MADCAM1 (MADCAM1-WT), MADCAM1^P270Q mutants (MADCAM1-P270Q), MADCAM1^D242N mutants (MADCAM1-D242N) or negative control (MADCAM1-NC) were treated with 100 μg/ml cycloheximide (CHX) and then were harvested for western blot at indicated time points. Western blot data are quantified by ImageJ software. D Macrophage migration assay by the supernatants from MADCAM1^WT, MADCAM1^MUT and control GC cells. E T cell migration assay by the supernatants from MADCAM1^WT, MADCAM1^MUT and control GC cells. F The chemokine profiles of corresponding supernatants detected by Human Chemokine Antibody Arrays. G Expression of CCL2 and IL8 in supernatant from MADCAM1^WT, MADCAM1^MUT and control GC cells. H Blockading CCL2 by anti-CCL2 antibody reduced migration of macrophages, especially in MADCAM1^D242N GC cells. I The p-Akt, CCL2 and p-mTOR was blocked by Akt inhibitor Perifosine, while the total levels of Akt were hardly changed. J The expression of p-mTOR and CCL2 was obviously suppressed after inhibition of Akt in MADCAM1^P270Q and MADCAM1^D242N GC cells. Bars, ±SD; *p < 0.05; **p < 0.01. ***p < 0.001.