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

Figure 5.

Figure 5

Neutrophil extracellular traps (NETs) contain selected enzymes from neutrophil cytoplasmic granules. Top panel shows human IgG staining (green), and bottom panel shows overlay of human IgG (green) and DNA staining (white). (A–D) Stainings of NETs from phorbol 12-myristate 13-acetate-activated neutrophils. DNA staining (white) shows NETs as elongated fibers. Bar represents 50 µm. (A) Staining with anti-myeloperoxidase (MPO)-IgG (green) shows NETs. (B) Anti-proteinase 3 (PR3)-antibodies do not stain NETs (green), but label cell bodies of netting neutrophils. (C) Anti-lactoferrin (LF) antibodies and (D) anti-neutrophil elastase (NE)-IgG stain NETs. (E–H) Staining of naïve neutrophils along with spontaneous NETs. DNA staining shows nuclei of neutrophils and spontaneously formed NETs. Bar represents 20 µm. (E) Anti-MPO-IgG stained spontaneously formed NET along with the cytoplasm of naïve cells. (F) Anti-PR3-IgG stained the cytoplasm of naïve cells but not spontaneously formed NET. (G) Anti-LF antibodies and (H) anti-NE antibodies stained the cytoplasm of naïve cells and spontaneously formed NET. Images are representative of at least three independent experiments.