Fig. 2

Detection of Sytox-stained NETs by DANA in rheumatoid arthritis subjects and controls. Neutrophils from rheumatoid arthritis (RA) subjects and controls were incubated for 2 and 4 h on coverslips, then fixed, stained with Sytox, mounted, and imaged at 200×. a. Two individuals independently counted the number of nuclei and NET-like structures by eye and then DANA was used to calculate the percent NETosis for all images (n = 78 images). The mean absolute differences with SEM for percent NETosis per image for DANA versus individual 1 by eye, DANA versus individual 2 by eye, and individual 1 versus individual 2 by eye were graphed. DANA was then used to quantify percent NETosis (b) and DNA area (c) with average and SEM graphed for RA versus control neutrophils (2 h RA, n = 24 subjects; 2 h control, n = 11 subjects; 4 h RA, n = 21 subjects; 4 h controls, n = 17 subjects; *p < 0.05)