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. 2020 Nov 9;10:19360. doi: 10.1038/s41598-020-76377-8

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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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Mouse treatment with aspirin and/or rivaroxaban can inhibit the number of bound platelets per leukocyte. Blood was collected from mice at 2 weeks after temporary ligation of the carotid artery. During the post-ligation period, mice were treated with aspirin (ASA), rivaroxaban (RIVA), ASA + RIVA, or saline (control). Using flow cytometry, platelet-leukocyte aggregates (PLAs) were measured in whole blood by dual staining with APC-αCD45 and DyLight 488-labelled αGPIbβ antibodies. Platelets were unstimulated or stimulated with 2-MeSADP (5 μM) or AYPGKF (30 μM). (A) Flow cytometric events were gated for CD45 fluorescence, and size anomalies were excluded by forward sideward scatter analysis. Shown is fluorescence in control blood from all leukocytes without (left of line) and with (right of line) bound platelets as defined by DyLight 488-labelled αGPIbβ fluorescence. (B) Calculated number of bound platelets per PLA. Data represented as mean ± SEM, n = 11 (control), 11 (ASA), 12 (RIVA) and 6 (ASA + RIVA) animals per group, *p < 0.05, **p < 0.01, ***p < 0.001, two-way ANOVA.