Vav2 in platelets.

(A) Human platelets were stimulated with CRP (5 μg/mL, 60 seconds) and lysis buffer added. Vav2 was immunoprecipitated from these samples with a rabbit polyclonal antibody to Vav2 and analyzed by immunoblotting with a second antibody to Vav2 (bottom panel). Immunoblots were then stripped and reprobed with monoclonal antiphosphotyrosine antibody 4G10 (top panel). (B) PLCγ2 was immunoprecipitated from control and Vav1^−/−Vav2^−/− mouse platelets stimulated with CRP at the indicated concentrations for 60 seconds. The resulting immunoblots were probed for phosphotyrosine (anti–p-Tyr), stripped, and reprobed for PLCγ2. (C) Platelets from wild-type, Vav1^−/−, Vav2^−/−, and Vav1^−/−Vav2^−/− mice, without EGTA, indomethacin, or apyrase were stimulated with CRP (0.2 μg/mL) or thrombin (0.03 U/mL) and the change in optical density indicative of aggregation monitored. One experiment representative of 3 is depicted.