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. 2021 Nov 30;144(24):1940–1954. doi: 10.1161/CIRCULATIONAHA.121.056414

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© 2021 The Authors.

Circulation is published on behalf of the American Heart Association, Inc., by Wolters Kluwer Health, Inc. This is an open access article under the terms of the Creative Commons Attribution Non-Commercial-NoDerivs License, which permits use, distribution, and reproduction in any medium, provided that the original work is properly cited, the use is noncommercial, and no modifications or adaptations are made.

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Figure 6. — Increased platelet activation, STAT signaling, and NETosis in human induced pluripotent stem cell–derived LNK(TT) cells. A, Surface P-selectin level on megakaryocytes (MMKs)/platelets with and without ADP (20 µmol/L) and thrombin (1 U/mL) or AYPGKF (50 µmol/L) stimulation from day 15 LNK(TT) and LNK(CC) cells (n=3). Unpaired t test. B, p-STAT5 level was analyzed in day 13 CD45⁺CD14^– LNK(TT) and LNK(CC) cells (n=3). Unpaired t test. C, MMKs/platelets activated with 1 nmol/L thrombin were incubated with LNK(TT) and LNK(CC) neutrophils for 4 hours. Representative images of NETosis. Green, Neutrophil elastase; red, citrullinated histone (H3Cit); blue, DAPI. Neutrophil extracellular trap (NET) numbers were normalized to neutrophil elastase positive cell numbers; 2-way analysis of variance. Scale bar, 50 µm. D, LNK(TT) and LNK(CC) neutrophils were stimulated with 5 µmol/L platelet-activating factor (PAF) for 3 hours and quantified for NETs (n=6); 2-way analysis of variance. Scale bar, 50 µm. Data are expressed as mean±SEM.