Figure 1.
Absence of ET formation by eosinophils. Eosinophils and neutrophils were preincubated with 10 μM DPI or 0.1% DMSO as vehicle control where indicated. To induce ET formation, eosinophils and neutrophils were stimulated with (A-C,E) PMA (100 ng/mL), (D) MSU crystals (200 μg/mL), or opsonized C. albicans for 4 hours at 37°C. (A-B,D) DNA release of eosinophils and neutrophils was measured in real time by Sytox Green Nucleic Acid Stain and (C) the area under the curve was calculated (mean + standard deviation , n = 6-7 for PMA vehicle condition, n = 6-7 for MSU crystals condition, n = 4-5 for C. albicans condition, n = 3-4 for DPI conditions). (E) ETs were visualized by staining for NE (green, 488), DNA (Hoechst, blue, 405), and myeloperoxidase (MPO, magenta, 633). Images were acquired using a Leica SP8 confocal microscope, original magnification ×400. Scale bar = 20 μm. Results are representative of 3 independent experiments. DMSO, dimethyl sulfoxide.

Absence of ET formation by eosinophils. Eosinophils and neutrophils were preincubated with 10 μM DPI or 0.1% DMSO as vehicle control where indicated. To induce ET formation, eosinophils and neutrophils were stimulated with (A-C,E) PMA (100 ng/mL), (D) MSU crystals (200 μg/mL), or opsonized C. albicans for 4 hours at 37°C. (A-B,D) DNA release of eosinophils and neutrophils was measured in real time by Sytox Green Nucleic Acid Stain and (C) the area under the curve was calculated (mean + standard deviation , n = 6-7 for PMA vehicle condition, n = 6-7 for MSU crystals condition, n = 4-5 for C. albicans condition, n = 3-4 for DPI conditions). (E) ETs were visualized by staining for NE (green, 488), DNA (Hoechst, blue, 405), and myeloperoxidase (MPO, magenta, 633). Images were acquired using a Leica SP8 confocal microscope, original magnification ×400. Scale bar = 20 μm. Results are representative of 3 independent experiments. DMSO, dimethyl sulfoxide.

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