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. 2016 May 20;36(3):e00333. doi: 10.1042/BSR20160031

Figure 5. NADPH-oxidase and elastase contribute to DFO-mediated NET formation.

Figure 5

Human blood-derived neutrophils were isolated by density gradient centrifugation and NET formation was visualized using the PL2–6 antibody against H2A–H2B–DNA complexes (green) in combination with DAPI to stain the nuclei (blue). (A) Cells were incubated for 3 h in media only or media containing 300 μM DFO or 25 nM PMA in the presence and absence of DPI (10 μg/ml). (B) Cells were incubated for 3 h in media only or media containing 300 μM DFO in the presence and absence of aprotinin (40 μg/ml). (C) Cells were incubated for 3 h in media only or media containing 300 μM DFO in the presence and absence of 0.1 mM elastase inhibitor N-(methoxysuccinyl)-Ala-Ala-Pro-Val-chloromethyl ketone. All graphs represent the mean ± S.E.M. of a minimum of 18 images derived from three independent experiments.