Fig. 4: Neutrophil-derived migrasomes activate platelets in vitro.
From: Neutrophil-derived migrasomes are an essential part of the coagulation system
a,b, Flow cytometry analyses of platelet activation. Platelets were isolated from neutrophil-depleted mouse blood and stimulated with PBS, thrombin (1 U ml−1) or nsNeuMig (nsNeuMig:platelet = 1:2). Platelet activation is indicated by CD62P (a) and platelet morphology is indicated by SSC and FSC (b). c, Platelets activated by thrombin or neutrophil-derived migrasomes were stained with APC–anti-CD41 (cyan), PE–anti-CD62P (purple) and AF488–anti-Ly6G (yellow) and imaged by three-dimensional Dragonfly confocal microscopy. Scale bars, 20 μm. d, Measurement of the diameter and area of platelets (Ctrl) and platelet aggregates induced by thrombin and nsNeuMig. n = 104 platelets for Ctrl and n = 106 and n = 138 platelet aggregates for thrombin and nsNeuMig, respectively. e, SEM images of platelets activated in vitro by thrombin or nsNeuMig. Yellow arrowheads indicate migrasomes coated with anti-Ly6G-conjugated magnetic beads. Cyan arrowheads indicate platelets. Scale bar for the left three panels, 2 μm. The migrasomes and platelets in the dashed box are enlarged on the right (scale bar, 1 μm). f, Measurement of platelet protrusion length. n = 61 platelets for Ctrl, n = 64 platelets for thrombin and n = 62 platelets for nsNeuMig. g, Flow cytometry analysis of platelet activation. Platelets were isolated from neutrophil-depleted mouse blood and stimulated with PBS, thrombin or neutrophil-derived migrasomes (nsNeuMig:PLT = 1:2, 1:10, 1:20, 1:50, 1:100 or 1:300). Platelet activation is indicated by CD62P. All statistical data are presented as means ± s.e.m. P values were calculated using a two-tailed, unpaired t-test. Source numerical data are available in Source Data Fig. 4.
