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. 2017 Apr 13;8:439. doi: 10.3389/fimmu.2017.00439

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Copyright © 2017 Panda, Krieger, Hopf, Renné, Haag, Röber, Conrad, Csernok and Fuchs.

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) or licensor 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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Comparison of anti-neutrophil cytoplasmic antibody (ANCA) stainings on ethanol-fixed neutrophils and NETotic neutrophils. Overlay of DNA staining (white) and human IgG staining (green). (A–C) Staining of NETotic neutrophils. (A) P-ANCA positive sera stain the nucleus of NETotic neutrophils. (B) C-ANCA positive sera stain the cytoplasm of NETotic cells. (C) Staining with autoantibody-free sera (IgG Ctrl) did not stain the cells. (D–F) Staining of ethanol-fixed naïve neutrophils with ANCA-positive sera. (D) P-ANCA positive sera stain the nucleus of ethanol-fixed naïve neutrophil. (E) C-ANCA positive sera stain the cytoplasm. (F) Staining with autoantibody-free sera (Ctrl) did not stain the cells. Images are representative of at least three independent experiments. Bar represents 50 µm.