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Figure 6 Changes in PAC-1 binding to activated platelets caused by adenosine diphosphate (ADP) receptor antagonists. To activate platelets, 25 µl of HEPES buffer containing ADP and epinephrine (400 µmol/l each) was mixed with 375 µl of platelet-rich plasma and incubated for 20 min. Fifty µl of fluorescein isothiocyanate-conjugated PAC-1 (25 µg/ml) was then added, and the fluorescence of individual platelets was measured 5, 15, 30, 45, and 60 min after the addition of 50 µl of HEPES buffer containing or not the P2Y12 antagonist AR-C69931MX (100 nmol/l). (Upper panel) Mean and SEM of the median fluorescence of 10,000 platelets in eight experiments. (Lower panels) Representative flow cytometric results at selected time points in one experiment without (solid lines) or with (dotted lines) the addition of AR-C69931MX.
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