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. 2021 Nov 25;139(8):1222–1233. doi: 10.1182/blood.2021013422

Figure 2.

Figure 2

RvTs reduce NETs in human peripheral blood. Human blood (500 μL) was incubated with RvT1, RvT2, RvT3, and RvT4 (10 nM each) individually, or with a combination of RvTs (RvT1-RvT4; 2.5 nM each), RvD2 (10 nM), LTB4 (10 nM), or vehicle control (0.01% ethanol v/v in saline) at 37°C for 15 minutes followed by the addition of PMA (1 μg/mL) for 4 hours. Aliquots of blood (50 μL) were added to Sytox Green (450 μL) and loaded onto the inlet reservoirs of the microfluidic NET devices. NETs captured within the micropost arrays were then imaged and quantified. (A) (Top panels) Representative images of fluorescent NETs using Sytox Green. Scale bars = 100 μm. (Bottom panels) Results are expressed as NET areas. The NET areas of PMA and PMA plus RvTs or RvD2 obtained from the same donors are connected by green lines. n = 4 (RvTs) or n = 5 (RvDs) separate donors. *P < .05, ***P < .001, paired two-tailed Student t test. (B) (Top panels) Representative images of fluorescent NETs using Sytox Orange. Scale bars = 100 μm. (Bottom panel) The NET areas of PMA and PMA plus LTB4 obtained from the same donors are connected by red lines. n = 3 separate donors. *P < .05, paired two-tailed Student t test. Details on imaging and quantification are given in the “Materials and methods”. (C) Results are percent reduction of NETs compared with PMA alone. Mean ± SEM. n = 3 to 5. *P < .05, **P < .01, vs LTB4.