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. 2018 May 8;9:968. doi: 10.3389/fimmu.2018.00968

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Copyright © 2018 Muñoz-Caro, Conejeros, Zhou, Pikhovych, Gärtner, Hermosilla, Kulke and Taubert.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Quantification of different types of neutrophil extracellular traps (NETs) induced by Dirofilaria immitis stages. Canine polymorphonuclear neutrophils (PMN) (n = 3, 2 × 10⁵) we exposed to microfilariae (MF; 50 larvae) and L3 (10 larvae) on poly-l-lysine pre-coated coverslips for 1 and 3 h. After fixation, immunofluorescence analysis was performed using Sytox Orange as DNA-marker along with anti-histone antibodies. The types of NETs (diffNETs, sprNETs, and aggNETs) were identified microscopically and counted.