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. 2021 Dec 20;22(24):13646. doi: 10.3390/ijms222413646

Figure 6.

Figure 6

Postpartum NET formation is triggered by circulating microparticles. (a) In vitro NET formation detected microscopically in a 3 h time course with SytoxGreen DNA binding dye after a 1 h pretreatment with equal amounts of microparticles isolated from pregnancy plasma and 48 h postpartum plasma and SytoxGreen fluorimetry of in vitro extracellular DNA release by control neutrophils (right panel) after 3 h treatment with pregnancy and post-delivery microparticles. (b) Measurement of ROS generation by control neutrophils in response to pregnancy and postpartum derived microparticles by DCFH-DA detection. (c) Fluorescent immunostainings for MPO (red), citH3 (green) and DNA (blue) and (d) morphometric assessment of NETs (MPO positive) and pro-NETotic (citH3 positive) neutrophils under the same experimental setup. Magnification: 20×; Scale bars: 50 μm. P: pregnancy; PD: post-delivery. Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 (one-way anova followed by bonferroni’s multiple comparison post-test). All experiments were performed at least 3 times with consistent results.