Expression and phosphorylation of SLAM, SAP, and EAT-2 in murine platelets. (A) Left panel: representative washed mouse platelet aggregation measurement showing inhibition of 4 μg/mL collagen-induced aggregation by the specific murine αIIbβ3 inhibitor ML00341819. Right panel: Western blot of resting or 4 μg/mL collagen-induced aggregated samples in the absence (lane 2) or presence of 5 μM (lane 3) or 10 μM (lane 4) ML00341819. Lysed platelets were immunoprecipitated with α-SLAM (9D-1) IgG. (B) Western blotting of washed wt, SAP–/–, and SLAM–/– mouse platelets resting (R) or aggregated (A) with 0.5 U/mL thrombin. Lysed platelets were immunoprecipitated with either nonspecific (lane 1) or α-SLAM (9D-1) IgGs (lanes 2-7). For both A and B, immunoblotting was performed with α-phosphotyrosine PY20 and 4G10, α-SLAM, α-SAP, α-EAT2 antibodies as indicated. Data shown are representative of experiments performed 3 times.
