Figure 2.
PAR4-AP-induced calcium mobilization (A) and platelet aggregation (B) in human platelets. (A) Washed human platelets (1 × 106 platelets/mL) were stimulated with 200 µM PAR4-AP in the absence or presence of increasing concentrations of 35. Calcium mobilization was decreased as the concentration of 35 was increased. Calcium was measured in real time using a C6 Accuri flow cytometer. The experiments were done in triplicate. (B) Platelet aggregation of human platelets (3 × 108 platelets/mL) was measured in real-time using a Chronolog Lumi-Aggregometer (model 700D) following addition of PAR4-AP. While 10 µM 35 did not inhibit platelet aggregation, 25 µM 35 inhibited 80% platelet aggregation in washed human platelets.