Selection of proteins tyrosine phosphorylated after CLEC-2 platelet stimulation. (A) Phosphotyrosine immunoprecipitation analysis of several proteins detected to be altered in the proteomic experiment: ADAP, Fer, Dok-2, Btk, and SHIP-1. (B) Tyrosine phosphorylation of a selection of proteins identified in the tyrosine phosphoproteome analysis was further validated using a combination of immunoprecipitations and Western blot. (i) Fer. (ii) Dok-2. (iii) ADAP. Proteins were separated on 4%-12% NuPAGE Bis-Tris gels before Western blotting. Densitometry graphs representing average band intensities: *P < .05. **P < .01. Basal indicates basal platelets; Rhod, platelets stimulated with rhodocytin (300nM, 5 minutes); IP, immunoprecipitation; and IB, immunoblot. Experiments were done at least 4 times.