The Impact of Various Reactive Oxygen Species on the Formation of Neutrophil Extracellular Traps
Figure 6
Release of NETs by neutrophils after exposure to various ROS inhibitors as measured by the SYTOXgreen assay. Freshly isolated human neutrophils were incubated with or without inhibitors (10 μM Dinitrophenol, 500 nM FCCP, 20 μM DPI, 38 μM Aroclor, 200 μM Dipyrone, and 200 μM Aminopyrine) for 30 min at 37°C. Subsequently, 5 μM SYTOXgreen was added to all samples, and formation of NETs was induced by 20 nM PMA. Samples without PMA were used as control. The NET production was detected for a period of 4 h at 37°C by measuring the fluorescence of DNA-bound SYTOXgreen. (a) Time kinetics of NET release (fluorescence intensity) of one representative experiment. (b) Quantification of NET release by calculation of the area under the curve (AUC). Data show mean ± SD from three independent experiments, *: as compared to the PMA-stimulated sample without inhibitor (Medium).
