Differential effect of secondary mediators on Dok-2 and ADAP tyrosine phosphorylation after CLEC-2 activation. Platelets were stimulated with rhodocytin in the presence (+) and absence (−) of inhibitors of secondary mediators (apyrase and indomethacin). (A) Dok-2. (B) ADAP. (i) Representative images. (ii) Densitometry graphs representing average band intensities and statistics. *P < .05. Proteins were separated on 4%-12% NuPAGE Bis-Tris gels before Western blotting. B indicates basal platelets; Rhod, platelets stimulated with rhodocytin (300nM, 5 minutes); IP, immunoprecipitation; and IB, immunoblot. Experiments were done at least 4 times.