Gads is tyrosine phosphorylated in response to CRP in platelets. (A) Close-up image of representative 2D gel images showing the Gads features. (B) Gads is tyrosine phosphorylated in response to CRP in human platelets, with maximum phosphorylation levels observed after 90 sec stimulation. Gads immunoprecipitated proteins were separated by 4-12% NuPAGE Bis-Tris gels and immunoblotted using anti-4G10 or anti-Gads antibodies. MS/MS analysis of the bands indicated by arrows, from gels run alongside the blots, allowed the identification of SLP-76 and Gads in the same signaling complex. Swiss-Prot accession numbers are shown in brackets before the protein names. B, basal platelets; CRP, platelets stimulated with CRP (10 μg/mL, 90 sec); IP, immunoprecipitation; IB, immuno blot; WCL, whole cell lysate. Images represent at least three independent experiments.