Fig 1: Genetic deficiency of platelet-specific FasL leads to reduced apoptosis post-AMI. (A) FasL exposure at the platelet surface of naïve mice and 6 h, 24 h, and 5 days post-AMI; n = 13. (B) Plasma levels of soluble FasL (sFasL) in naïve mice and 6 h, 24 h and 5 days post-AMI; n = 5–7. (C,D) mRNA analysis of pro-apoptotic Bax (C) and anti-apoptotic Bcl2 (D) genes in the LV of FasLfl/fl-Pf4-Cre− and FasLfl/fl-Pf4-Cre+ naïve mice and 24 h post-AMI; n = 5. (E) Plasma levels of sFasL in FasLfl/fl-Pf4-Cre− and FasLfl/fl-Pf4-Cre+ naïve mice and 24 h post-AMI; n = 4–9. (F) Quantification (left panel) and representative images (right panel) of caspase-3 positive cells in the heart of FasLfl/fl-Pf4-Cre- and FasLfl/fl-Pf4-Cre+ mice 5 days after AMI; n = 4–5. (G) Echocardiographic analysis of cardiac function by determination of ejection fraction (baseline vs. 24 h after I/R) of FasLfl/fl-Pf4-Cre− and FasLfl/fl-Pf4-Cre+ mice; n = 12–13. (H) Quantitative analysis (left panel) of infarct size as the percentage of the area at risk (% Inf/AAR) with representative images (right panel) of FasLfl/fl-Pf4-Cre− and FasLfl/fl-Pf4-Cre+ mice 24 h post-AMI. Blue = healthy tissue, red = the area at risk (AAR), white = infarcted area (INF); n = 7. Scale bar = 50 µm. White bars = FasLfl/fl-Pf4-Cre-; green bars = FasLfl/fl-Pf4-Cre+ Data are presented as means ± SEM. Statistical analyses were carried out by two-way ANOVA followed by Sidak post hoc (A–E,G) or by two-tailed unpaired Student’s t-test (F,H). * p < 0.05, ** p < 0.01, *** p < 0.001.
Fig 2: Increased adhesion of platelets to immobilized FasR under hypoxic conditions. (A) Representative images and (B) Quantification of adherent resting and ADP [10 µM]-stimulated human platelets to immobilized human recombinant FasR protein (50 µg/mL) under normoxic (21% O2) or hypoxic (2% O2) conditions (n = 4–6). (C) Externalization of FasL on the surface human platelet determined by flow cytometry (n = 5). (D) Annexin V binding to resting and CRP [5 µg/mL]-stimulated human platelets under normoxic (21% O2) or hypoxic (2% O2) conditions (n = 5–6) determined by flow cytometry. (E) Soluble (s)FasL plasma level (in ng/mL) from patients with coronary heart disease (CHD) compared to ST-elevation myocardial infarction (STEMI)-patients (n = 11–14). Scale bar = 50 µm (missing). Data are presented as means ± SEM. Statistical analyses were carried out by (B) two-way ANOVA followed by Sidak post hoc; (C,E) by two-tailed unpaired Student’s t-test, and (D) multiple t-test. * p < 0.05, ** p < 0.01.
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