Dok-1 is tyrosine phosphorylated and associates with RasGAP, Nck, and Crk-L after SDF-1α/CXCL12 stimulation. (A) Jurkat cells were left unstimulated (–) or stimulated with SDF-1α/CXCL12 (100 ng/mL) for 5 minutes. Total cell lysates (TCLs) were immunoprecipitated with anti–Dok-1 Ab. Immunoprecipitates (IPs) were immunoblotted with antiphosphotyrosine antibody (upper panel) or anti–Dok-1 Ab (middle panel). As control, IPs were made with control IgG, and immunoblotting was performed with antiphosphotyrosine antibody (lower panel). (B) Immunoprecipitates were immunoblotted with anti-RasGAP, Nck, and Crk-L antibodies. (C) As a control for panel B, IPs with control IgG Ab were immunoblotted with anti-RasGAP, anti-Nck, or anti–Crk-L Abs. (A-C) Results are representative of at least 3 experiments. WB indicates Western blot.