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. 2020 Nov 16;10:19876. doi: 10.1038/s41598-020-76428-0

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© The Author(s) 2020

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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Comparison of EGFP expression in immune cells of the brain. (A) Brain microglia (CD11^bCD45^low) from wt and BAC transgenic mice were analysed by flow cytometry for endogenous EGFP expression and cell surface P2X7 expression using a monoclonal antibody (clone RH23A44). (B) Immunofluorescence staining of Iba1 and EGFP in the third ventricle of the P2X7-EGFP mouse. Scale bar: 25 μm. (C) Comparison of perivascular macrophage staining in sEGFP and P2X7-EGFP mice. Anti-collagen IV antibody was used to stain blood vessels and CD206 was used as a marker for perivascular macrophages. Lower panel: detailed images from the sEGFP mouse showing a lack of overlap between EGFP and CD206 staining. Scale bar:10 µm, DAPI staining in blue. (D) Peritoneal macrophages (CD11b⁺FceR1^–) and mast cells (CD11b^–FceR1⁺) from wt and BAC transgenic mice were analysed by flow cytometry for endogenous EGFP expression and cell surface P2X7 expression. (E) CD4⁺ T cells from the spleen wt and BAC transgenic mice were analysed by flow cytometry for endogenous EGFP expression and cell surface P2X7 expression. EGFP and P2X7 expression levels were compared on helper T cells (CD4⁺CD25^–) and regulatory T cells (CD4⁺CD25⁺).