Inhibition of NETosis Pathways

Specific inhibitors described previously were used for blockage of NET formation and have been described previously^{23 (link),34 (link),40 (link),46 (link),90 (link)–94 (link)}. The following inhibitors were used: the NE inhibitor Suc-Ala-Ala-Pro-Val chloromethyl ketone (CMK; 1 mM final concentration, Sigma), NADPH oxidase (NOX) inhibitor diphenylene iodonium (DPI; 10 μM final concentration, Sigma), the MPO inhibitor 4-Aminobenzoic acid hydrazide (ABAH; 100 μM final concentration, Sigma), TLR4 signaling inhibitor CLI-095 (1 μg/mL final concentration; InvivoGen), the TLR4 signaling inhibitor polymixin B (100 μg/mL final concentration; InvivoGen, San Diego, CA), the PKC inhibitor Bisindolylmaleimide I (310 nM final concentration, Cayman Chemical, Ann Arbor, MI), and the ROS scavenger inhibitor ascorbic acid (Vitamin C) (200 μM final concentration, Cayman Chemical, Ann Arbor, MI). Cells were pre-incubated with inhibitors for 30 min at 37 °C prior to stimulation as described above^{23 (link),34 (link)}.