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

Figure 1. DFO induces NET formation in human neutrophils.

Figure 1

Human blood-derived neutrophils were isolated by density gradient centrifugation and treated with DFO in the presence and absence of excessive iron ions. Formation of NETs was visualized using the PL2–6 antibody against H2A–H2B–DNA complexes (green) in combination with DAPI to stain the nuclei (blue). (A) Increased NET formation was detected for neutrophils stimulated for 3 h with 300 μM DFO or 25 nM PMA as positive control. (B) Representative fluorescent micrographs of human neutrophils incubated in media only, media containing 300 μM DFO or 25 nM PMA as positive control representing the results of the column bar graph in (A). (C and D) Addition of divalent or trivalent iron ions (250 μM) abolishes the NET-induction effect by DFO. The graphs represent the mean ± S.E.M. of 60 images from 10 independent experiments (A) or 18 images derived from three independent experiments (C and D).