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. 2022 Mar 18;13:1481. doi: 10.1038/s41467-022-29151-5

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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 Venn diagram analysis of upregulated genes in G600, Over-Expression-S100a9 in B477 cells (B477-OE-S100a9), and down-regulated genes in G600 cells expressing sgS100A9 (G600-sgS100a9) (n = 3 biological replicates/group). b KEGG Pathway analysis with 453 common (p value was calculated by hypergeometric test with KOBAS 3.0 website). c Top 20 differentially expressed genes by comparison of the gene expression profiles of four different group cells, including B477-Ctr (B1-3), B477-OES100a9 (P1-3), G600-Ctr (G1-3), and G600-sgS100a9 (G10-1-3) (the heatmap was drawn by Morpheus website and clustered by One minus Pearson Correlation). d, e Expressions of S100a9 and Cxcl12 at mRNA level by qPCR (d) at protein levels by western blotting (e) from four group cells in c. f Protein levels of S100a9, Cxcl12 and pStat3 in OE-S100a9-EMT6 cells, sgS100a9/OE-S100a9-EMT6, and sgCxcl12/OE-S100a9-EMT6 cells. g Protein levels of S100a9, Cxcl12 and pStat3 in OE-S100a9-EMT6 and G600 cells without or with S100A9 inhibitor, Tasquinimod (Tas-50 μM) by western blotting. h Protein levels of S100a9, Cxcl12, and pStat3 in WT (B477) cells with the expression of OE-S100a9, OE-S100a9/sgCxcl12, or OE-Cxcl12 by western blotting. i, k Protein levels of Cxcl12 in B477 (i) cells with OE-S100a9 and in G600 (k) cells expressing sgS100a9 at different time courses (0–48 h) and different amounts (0–4 μg) by western blotting. j, l Protein levels of S100a9 in B477 (j) cells with OE-Cxcl12 and in G600 (l) cells expressing sgCxcl12 at different time courses (0–48 h) and different amounts (0–4 μg) by western blotting. m Protein levels of S100a9, Cxcl12, and pStat3 in G600 cells after treating with Tas and FPS-ZM1, inhibitor for RAGE receptors on cell membranes by western blotting. n Protein levels of S100a9, Cxcl12, and pStat3 in B477 cells with the expression of shBrca1 and MT G600 cells with over-expression of Brca1 (OE-Brca1). o A diagram summarizing the relationship of Brca1, S100a9, Cxcl12 and pStat3 in Brca1-MT mammary epithelial cells, and their potential interaction with the surrounding immune microenvironment, which remains elusive. The data are expressed as means ± SD (d) and P values determined by one-way ANOVA followed by Tukey’s multiple comparisons. The experiments were independently repeated three times with similar results (d–n). Source data are provided as a Source data file.