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Platelet aggregation and ATP release
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The platelet aggregation assay was performed as previously described [19]. Platelets were resuspended in HEPES-buffered Tyrode’s solution. The final platelet concentration was adjusted to 300 × 109/L, and the platelets were transferred to an aggregation tube for preincubation with different concentrations of NEFAs and CaCl2 (1 mM) at 37 °C for 5 min. Aggregation tube transmittance was measured using a platelet aggregation instrument for quantification. Platelet secretion was monitored by measuring ATP release using the Chrono-lume reagent (Chrono-Log).
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