1. Fig. 4. Neutrophil aggregation is selectively induced by PS+ platelets.
Neutrophil aggregation is selectively induced by PS+ platelets. (A to C) Mice were administered indicated fluorescence probes (A and C) or Alexa 488–annexin V (PS+ platelets) and DyLight 647–anti-GPIbβ antibodies (platelets) (B) before needle puncture of mesenteric veins, followed by local microinjection of either ADP or Thr/CRP. (A) Representative confocal images depict neutrophil aggregate formation and detachment (red) from ADP or Thr/CRP thrombi (green) ~30 min after agonist injection. (B) Annexin V binding to thrombi (PS+ platelets) at the indicated times after agonist injection (n = 3). (C) Confocal images depicting PS+ platelets (green/yellow) within detaching neutrophil aggregates (red) from Thr/CRP-stimulated thrombi. (D and E) Confocal images depicting neutrophil aggregates (red) anchored by PS+ platelets (green/blue, cyan) in mesenteric veins, as demonstrated by channel overlays (white and cyan in magnified images, respectively) (D), and in pulmonary vasculature (dotted line) (E) after gut I/R injury. (F) Confocal images depicting PS+ platelet formation (green/blue, cyan) in intestinal microvasculature during gut I/R injury. (G and H) Percent (%) of intestinal vessels containing PS+ platelets (G) and the percent of platelets in a PS+ state in the intestinal microvasculature (H) after I/R injury [I/R, n = 4 (G); sham, n = 3 (G); I/R, n = 5 (H)]. (I) Confocal image depicting PS+ platelets [green/blue, cyan (arrows)] adherent to pulmonary vasculature (dotted line) after gut I/R injury. (J) Confocal images depict occlusive neutrophil aggregates (green) and colocalized fibrin (red) and platelets (blue) in the pulmonary vasculature 2 hours after gut I/R injury (n = 5). Scale bars, 50 μm (D), 20 μm (E), 100 μm (F), 10 μm (I), and 50 μm (J). Error bars represent means ± SEM. *P < 0.05; *P < 0.01; ***P < 0.005; ****P <
Yuping Yuan et al., Sci Transl Med 2017;9:eaam5861
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