Kinetics of NET component presence on liver vascular endothelium and in blood followed for 365 days (1 year) after induction of endotoxemia. Histone H2A.X (A), NE (B), and extDNA (C) were followed by IVM on liver endothelium and by enzyme-linked immunosorbent assays (ELISAs) in blood serum. Additionally, NE proteolytic activity was measured in vivo. Endotoxemia was induced by LPS (IP; 1 mg/kg b.w.), and the parameters were estimated at given time points (hours, days, and months) within the following year. Data concerning NET components attached to the endothelium of liver sinusoids are marked with solid lines: histones H2A.X (red), NE (violet), NE activity (orange), and extDNA (green). The presence of the above components in blood serum is labeled with dashed black lines in panels A-C. (D) Representative images (IVM) of NETs for data presented in panels A-C (images for all time points are presented in supplemental Figures 8 and 9). The NET components were detected with AF568–anti-H2A.X antibody (red), AF647–anti-NE antibody (violet), and SYTOX Green labeling extDNA (bright green). The colocalization of NET components was visualized by overlaying the images from individual channels. The data in the graphs are expressed as means ± standard deviation of at least 3 fields of view. Statistically significant differences according to 1-way analysis of variance (ANOVA) are designated by letters, in which the same letter indicates no differences between groups, and different letters indicate statistical differences (Bonferroni post hoc; n = 3-4).