NET Visualization
NET visualization was performed according to previous protocols. The
neutrophils used for the plate reader assay were fixed with 4% PFA in
PBS buffer for 30 min and permeabilized with 0.1% Triton X-100 for 30
min. Then, blocking was performed with 10% normal goat serum for 30 min
at RT. The cells were stained using an anti-myeloperoxidase antibody
(Abcam ab25989) and an anti-histone H3 (citrulline2+8+17) antibody
(Abcam ab5103). A goat anti-mouse IgG H&L-Alexa Fluor® 647 antibody
(Abcam, ab150115) and a goat anti-rabbit IgG H&L-Alexa Fluor® 488
antibody (Abcam, ab150077) were utilized as secondary antibodies. The
DNA was already stained with SYTOX Green after permeabilization. The
images were captured using a Leica DMi8 inverted microscope.