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. 2024 Jan 20;31:10. doi: 10.1186/s12929-023-00991-7

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© The Author(s) 2024

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/. 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 in a credit line to the data.

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Fig. 6 — PTX3 contributes to M2 polarization through activation of CREB1 signaling. A RNA sequencing was performed in unstimulated THP-1 macrophages and PTX3-stimulated THP-1 macrophages. The top 200 upregulated and downregulated genes were selected for Connectivity Map analysis. Forward (overexpression) and reverse (knockdown) perturbation candidate genes were identified. B Western blot analysis of p-CREB1 and CREB1 in THP-1 macrophages treated with or without 50 ng/ml PTX3. C Western blot analysis of p-CREB1 and CREB1 in BMDM streated with or without 100 ng/ml PTX3. D The protein level and E mRNA level of CEBPB in THP-1 macrophages treated with or without 50 ng/ml PTX3. F, G The protein levels of p-CREB1, CREB1, and CEBPB in THP-1 macrophages treated with 100 ng/ml PTX3 plus either 500 ng/ml IgG1κ or WHC-001 (αPTX3Ab). H Western blot analysis of p-CREB1 and CREB1 in HEK293 cells transiently transfected with pcDNA3.1, CREB1-wt, or CREB1-S133A. I Illustration of the luciferase promoter constructs of CEBPB (− 890/ + 13), VEGF (− 1347/− 46), and ARG1 (− 919/ + 30). The CREB1 binding site is indicated in red and the CEBPB binding site is indicated in black. The promoter activity of the pGL3-CEBPB (− 890/ + 13), pGL3-VEGF (− 1347/− 46), and pGL3-ARG1 (− 919/ + 30) constructs in HEK293 cells cotransfected with pcDNA3.1, CREB1-wt, or CREB1-S133A is shown. J The promoter activity of pGL3-CEBPB (− 890/ + 13) in THP-1 macrophages treated with or without 50 ng/ml PTX3. K The promoter activity of pGL3-CEBPB (− 890/ + 13) in THP-1 macrophages cotransfected with CREB1-wt or CREB1-S133A following treatment with or without 50 ng/ml PTX3. L Schematic model showing that PTX3 contributes to stroma-mediated immunosuppression by promoting M2-like macrophage polarization through activation of the CEBPB/CREB1 axis in colon cancer. p values were calculated by two-tailed paired Student’s t test or one-way ANOVA. ns represents nonsignificant difference, *represents a p value of < 0.05, **represents a p value of < 0.01, and ***represents a p value of < 0.001. Values on the plots are presented as the means ± SEMs. Data are combined from 3 to 5 independent experiments