The Impact of Various Reactive Oxygen Species on the Formation of Neutrophil Extracellular Traps
Figure 2
Inhibitors of various steps of ROS production are not toxic and do not induce apoptosis in human neutrophils. Freshly isolated human neutrophils were incubated with or without inhibitors (10 μM Rotenone, 10 μM Dinitrophenol, 500 nM FCCP, 20 μM DPI, 38 μM Aroclor, 10 μM DETC, 200 μM Dipyrone, and 200 μM Aminopyrine) for 4 h at 37°C. DMSO and methanol were used as solvent controls for Rotenone and Aroclor. Apoptosis and cell death were assessed by flow cytometry using stainings with Annexin V-FITC and propidium iodide, respectively. (a) The effect of various inhibitors on apoptosis and necrosis of neutrophils in vitro. Data are shown as mean from 3 independent experiments. (b) Representative dot plot of a flow cytometry analysis of human neutrophils cultured without inhibitor. The -axis shows the Annexin V-FITC fluorescence, at the -axis the propidium iodide fluorescence is indicated. Events in the lower right quadrante (1.08%) represent the apoptotic cells whereas events in the upper quadrants represent the necrotic population.
