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. 2018 May 15;9:966. doi: 10.3389/fmicb.2018.00966

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Copyright © 2018 de Buhr, Bonilla, Jimenez-Soto, von Köckritz-Blickwede and Dolz.

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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T. cruzi induces ETs in didelphine granulocytes in vitro. Formation of ETs after infection of purified didelphine granulocytes with T. cruzi at MOI of 5 after 60 min (A) and 180 min (B) was analyzed. PBS was used as negative control and the cholesterol-depleting cyclodextrine (CD) as positive ET inducer. After infection ETs were visualized by immunofluorescence microscopy (ETs [DNA/histone-1-complexes] = green, Hoechst [DNA] = blue). Per sample at least 6 pictures were taken at predefined positions, and the number of granulocytes and the ET-releasing granulocytes were determined. (C) Representative pictures after 60 min of stimulation are shown. The isotype control was negative for ETs. Graphs in A and B show the mean ± SD of 4 independent experiments with 24 analyzed pictures. Statistical differences were tested by unpaired one-tailed Students t-Test (^****p < 0.0001).