OxLDL induces phosphorylation of Vav1 and Vav3 in platelets. Washed human platelets (2 × 108/mL) containing 2mM CaCl2 and 1mM MgCl2 were incubated with 50 μg/mL native LDL or oxLDL for 30 minutes (A) or 50 μg/mL oxLDL over varying time points (B,D) or various concentrations of oxLDL for 30 minutes (C) and then lysed. Vav1 (A-C) or Vav3 (D) was precipitated by specific Abs and precipitates were analyzed by immunoblot with 4G10 anti-phosphotyrosine Ab. The membranes were then stripped and reprobed with Abs to the total relevant proteins to normalize the protein loaded. Results are representative of at least 3 independent experiments from different donors. nLDL indicates native LDL; and oxLDL, oxidized LDL.