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Platelets were stimulated by collagen (1 g/mL) for the appropriate time at 37 C with stirring in Chrono-Log aggregometer and reactions were terminated by addition of equal volumes of chilled 2 NP-40 lysis buffer (100 mM Tris-HCL pH 7.4, 300 mM NaCl, 2 mM NaF, 2% NP-40, 2 mM EDTA, and protease and phosphatase inhibitor solution) on ice for 15 min