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. 2023 Aug 6;13:12740. doi: 10.1038/s41598-023-39904-x

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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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mRNA expression levels of cannabinoid receptors in various macrophage subtypes. (A–C) Human monocyte-derived macrophages (M0) were classified into three subtypes: M1 (CD80⁺CD86⁺; A), M2a (CD206⁺; B), and M2c (CD163⁺MerTK⁺; C). Surface expression of molecular markers was analyzed by flow cytometry, and representative histograms from three independent experiments are shown. Dotted line: isotype control. Solid line: antigen-specific antibodies. (D–F) mRNA expression of cannabinoid receptors GPR55 (D), CNR1 (E), and CNR2 (F) was quantified by RT-qPCR. Bars represent mean ± SEM from five independent experiments. Data were analyzed by one-way ANOVA and Tukey’s multiple comparison test (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001). (G) M0 and polarized macrophages (M1, M2a, M2c) were stained with DIM21 antibody, which recognizes PtdGlc-enriched microdomain, followed by Alexa Fluor 488-conjugated secondary antibody. Cells were analyzed by flow cytometry, and histograms are shown. Dotted line, isotype control. Solid line, DIM21.