FIG. 7.

An active Src kinase is obligatory for the interaction between Wrch1 and Pyk2. (A) The interaction between Myc-tagged Wrch1 or Wrch1 mutants and KD Pyk2 (Pyk2/KD) was assessed by transient transfections into HEK293T cells. The presence of Pyk2/KD in Myc immunoprecipitates (IP) was determined by immunoblotting (IB). wt, wild type. (B) The necessity of the presence of an active Src kinase for the Wrch1 and Pyk2 interaction was assessed by transient transfections of Myc-tagged Wrch1 or Wrch1 mutants and Pyk2 into HEK293T cells. The cells were treated with the Src inhibitor SU6656 (2 μM) for 1 h prior to cell lyses. No Pyk2 was detected in Myc immunoprecipitates, as determined by immunoblotting. (C) The necessity of the presence of the Src family kinase for the Wrch1-and-Pyk2 interaction was further assessed by transient transfections of Myc-tagged Wrch1 or Wrch1 mutants and Pyk2 into SYF^−/− MEFs. No Pyk2 could be detected in Myc immunoprecipitates, as determined by immunoblotting.