CLP36 associates in vivo with α-actinin in platelets.

(A) Coimmunoprecipitation of α-actinin in anti-CLP36 immunoprecipitates. Anti-CLP36 antibodies from 2 different rabbits (designated 1 and 2) were used. (B) No coimmunoprecipitation of CLP36 with the focal contact protein vinculin. The anti-CLP36 immunoprecipitates were resolved by SDS-PAGE, blotted, and probed with anti-CLP36, anti–α-actinin, or antivinculin antibody. Load indicates platelet lysate before immunoprecipitation corresponding to 7% of lysate used for immunoprecipitation. The experiment is representative for 10 independent experiments. (C) Interaction between CLP36 and platelet α-actinin as detected by the blot overlay assay. Platelet proteins were blotted, and the blot was probed with anti–α-actinin antibody (left) or with biotinylated-CLP36 (right).