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. 2020 Oct 22;10:18061. doi: 10.1038/s41598-020-73005-3

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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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Circulating extracellular vesicles (EVs) do not externalize PS and consequently do not support thrombin generation. (A) The procoagulant properties of 0–16 µg/ml circulating EVs (PFP) or stimulated PDEVs+ circulating EVs (PRP) were assessed using CAT in the presence of 1 pM TF. Dose–response curves representing peak thrombin generation are plotted against vesicle concentration. Results presented as Mean ± SD, n = 4. (B) Phosphatidylserine exposure was determined by Annexin V binding to PFP and PRP EVs and quantified through fluorescence flow cytometry. Due to low Annexin V binding, PFP EVs do not carry negatively charged phospholipids and consequently cannot support thrombin generation on its surface. Results presented as Mean ± SD, n = 8.