Fig 2. NET induction by Leishmania major is independent of NADPH oxidase activity.
(A) BMN (5 x 105 cells/ well) of B6 wild-type (WT; gray bars) or gp91phox-/- (hatched bars) mice were either left untreated (Nil) or incubated with increasing numbers of L. major promastigotes (1:1, 1:5) and NETs measured as the release of extracellular dsDNA and presented as fold increase over control 4 h post-stimulation. (B) ROS generation of bone-marrow-derived neutrophils of wild-type (black lines) or gp91phox-/- (purple lines) mice were measured every 5 min for a period of 90 min in the presence (filled lines) or absence of PMA (dashed lines). (C) BMN (5 x 105 cells/ well) of B/c (blue bars) or B6 (purple bars) mice were either left untreated (Nil) or incubated with an increasing ratio of L. major promastigotes (MOI 1 and 5) or PMA (100 nM) and (D) ROS generation was measured during 2 h with DHR123 probe. (E) Quantification of NET-DNA release 1 h post-stimulation with L. major promastigotes (MOI 1 and 5) of BMN from wild type (purple bars) and elastase knockout (green bars) mice. Results are mean±SEM of n = 4–6. Difference (#) between indicated bars were considered significant when p<0.05.