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. 2023 Jan 17;13:927. doi: 10.1038/s41598-023-28073-6

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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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VIP antagonist promoted M1 polarization and macrophage phagocytosis of CT26 in CT26-CM-incubated RAW264.7 cells. VIP concentration in whole-cell homogenate and supernatant collected from CT26 and RAW264.7 cells (A); n = 4–6/group. The effect of VIP hybrid (VIPhyb, a VIP antagonist) at 1 and 3 µM on mRNA expression of M1 macrophages (B), M2 macrophages (C), and immune checkpoint (D) markers in RAW264.7 cells at day 4 after incubation with CT26-CM. PBS was used as vehicle; n = 7–9/group. Representative FACS histograms and flow cytometric analysis of engulfed CT26 cells of CT26-CM-incubated RAW264.7 cells treated with vehicle (PBS) or VIPhyb at 3 µM (E). Gating strategies for phagocytosis of PKH-26-labeled CT26 in RAW264.7 cells are shown in Supplemental Fig. S2; n = 6/group. Data from each treatment were compared with data from the control; *p < 0.05 vs vehicle.