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. 2022 Dec 9;12:21313. doi: 10.1038/s41598-022-25176-4

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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 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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Nlrp3 deficiency reduces membrane expression of P-selectin on the inflamed endothelial cells, while circulating leukocyte adhesion receptors expression is not altered. Neutrophils and monocytes in peripheral blood were collected from Nlrp3^+/+ and Nlrp3^−/− mice 4 h after injection of thioglycollate broth (n = 5:6). (A) Surface expressions of integrins; integrin β2 (ITGB2, CD18), integrin αL (ITGAL, CD11a), integrin αM (ITGAM, CD11b), and P‐selectin glycoprotein ligand 1 (PSGL-1, CD162) were analyzed by flow cytometry. The relative MFI was calculated as a relative value to an average MFI of Nlrp3^+/+. (B-F) Endothelial cells were isolated from the mesentery of Nlrp3^+/+ and Nlrp3^−/− mice 4 h after injection with thioglycollate (Thioglycollate, closed circles, n = 9:7) or PBS (Control, open circles, n = 5:6). Surface P-selectin expression was analyzed by flow cytometry. (B–E) Gating strategy for isolated endothelial cells. Endothelial cells were defined as negative for CD45.2 and positive for CD31; (B) ungated cells and singlets, (C) live cells as negative for DAPI, (D) CD45.2 negative and (E) CD31 positive. (F) P-selectin expression on gated endothelial cells. Left: percentage of P-selectin positive endothelial cells. Right: mean fluorescent intensity (MFI) of P-selectin.