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Primary blood-derived neutrophils were isolated from fresh blood by density gradient centrifugation using Polymorphprep™ (Progen Biotechnik, Heidelberg, Germany), as described previously [18]. For in vitro NET formation assays, the neutrophils were seeded on poly-L-lysine-coated glass slides in 24-well plates at a concentration of 5 × 105 cells per well (250 μl) in RPMI 1640 medium (Thermo Fisher Scientific Inc., Waltham, MA) at 37 °C and 5% CO2, and NET formation was analyzed after incubation for 2 and 4 h, respectively.
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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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