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Platelet secretion was determined by measuring ATP release using luciferin/luciferase reagent (Chrono-Lume, Chrono-log Corporation, USA). Briefly, 90 μl of platelets (1 × 108/mL) in Tyrodes-HEPES buffer (with calcium) were incubated with 10 μl of NETs with gentle shake at 37 °C for 1 and 10 min before adding 5 μl of Chrono-Lume reagent. The luminescence was measured using Enspire Multimode Plate Reader (PerkinElmer, USA). Where anti-platelet drugs were used, platelets were pre-incubated with the drugs for 15 min at 37 °C before incubating with NETs.
Copyright and License information: The Author(s). ©2018
Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
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