Neutrophil Extracellular Traps and Microcrystals

Figure 3

PMNs release extracellular DNA in response to Alhydrogel in vitro. Human PMNs seeded on a 96-well black microplate were incubated for 30 minutes in the presence or absence of 10 μM DPI prior stimulation with increasing doses of commercially available Alhydrogel (InvivoGen, cat#: vac-alu-50) or 100 nM PMA. Increase in fluorescence due to extracellular DNA (ecDNA) release was measured in presence of 10 μM Sytox Orange DNA-binding dye for 5 hours with a microplate fluorimeter. DNA release is presented as either relative fluorescence units (RFU) or percentage of maximal DNA released achieved by saponin treatment [4, 20]. (a) Summary of three independent experiments using PMNs obtained from independent human donors. Mean +/− SEM. (b) Representative kinetics of fluorescence results (). Ut, untreated; PMA, phorbol myristate acetate.

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