Fig. 5.
De novo phosphorylation of CDCK5R1 after silencing of CDCP1. a Loss of CDK5R1-CDK5 complex formation in suspended CDCPl-silenced cells. Reciprocal immunoprecipitations of CDK5 or CDK5R1. WCL whole cell lysate, CAB IgG control, IP immunopre-cipitate. b ITGB1-CDK5 complex formation. Western blot of ITGB1/β1-integrin immunoprecipitation (ITGB1 IP) from DU145 cells with and without silencing of CDCP1 and probed for CDCP1, CDK5, and CDK5R1. c CDK5R1 immunoprecipitation of suspended DU145/ shCDCPl. CDK5R1 complexes from DU145/shControl (Cnt) or shCDCPl(shl) were probed with the 4G10 antibody. Western blot membranes were re-probed with antibodies reactive to PKCδ, SRC and CDK5R1. d Regulation of ITGB1/β1-integrin inside-out activity by c-SRC. DU145/shCDCP1 cells were treated with 10 μM Sar-acatinib or buffer and suspended for 3 h. Western blot probed with antibodies reactive to pTalin-S425, total Talin (Talin), HUTS-4 (ITGB1*), or total ITGB1/β1-integrin (ITGB1). e-f Re-adhesion of suspended DU145/shCDCP1 to fibronectin (e). Image of plate after adhesion of DU145/shCDCP1 cells with and without treatment with Vanadate treatment and quantification of adherent cell numbers (f). Western blot tyrosine phosphorylated proteins visualized with the 4G10 antibody. *P <0.05 and **P <0.01