Figure 6.

Effect of mtDAMP pre-treatment on neutrophil NET generation. (A) Serum concentrations of the mitochondrial-derived N-formylated peptide ND6 in peripheral blood samples acquired from healthy controls (HC) and trauma patients. Number of samples are indicated below each time-point. ^***p < 0.0001 vs. HC. (B) NET production by PMA stimulated neutrophils pre-treated with 40 or 100 μg/ml mtDAMPs was assessed by measuring DNA content of cell-free supernatants (n = 5). ^*p < 0.01 vs. Vehicle. (C) MtDAMP-induced inhibition of NET generation was confirmed by fluorescence microscopy (n = 5; top panel x20 magnification; bottom panel x40 magnification). (D) ROS generation by neutrophils pre-treated with 100 μg/ml mtDAMPs or vehicle control was measured in response to stimulation with 25 nM PMA using luminol-based chemiluminescence (n = 10). Data are presented as area under the curve (AUC) and are mean ± SEM. ^***p < 0.0001 vs. PMA.