Cdo and Abl interact through their proline-rich motif and SH3 domain, respectively. (A) Yeast cells transformed with the indicated vectors were tested for expression of β-Gal, indicative of interaction. AD, Gal4 activation domain; BD, Gal4 DNA-binding domain. (B) Lysates of 293T cells transiently transfected with GST-Cdo intracellular region (GST-Cdointra), Abl, AblΔSH2, AblΔSH3, or control (−) expression vectors, as indicated, were pulled down with glutathione-Sepharose beads and blotted with an antibody to Abl. Straight lysates were also Western blotted (WB) with antibodies to the Cdo intracellular region or Abl. (C) Lysates of 293T cells transiently transfected with Cdo, a deletion mutant form of Cdo, Abl, or control (−) expression vectors, as indicated, were immunoprecipitated (IP) with Abl antibodies and then Western blotted with an antibody to Cdo. (D) Lysates of 293T cells transiently transfected with GST-Cdointra, GST-CdointraP/A, Abl, or control (−) expression vectors, as indicated, were pulled down with glutathione-Sepharose beads and blotted with antibodies to Abl or Cdo. Straight lysates were also Western blotted with antibodies to Abl or Cdo. (E) Lysates of 293T cells transiently transfected with GST-Cdointra, GST-CdointraP/A, S epitope-tagged JLP (JLP-S), or control (−) expression vectors, as indicated, were pulled down with glutathione-Sepharose beads and blotted with antibodies to the S epitope or Cdo. Straight lysates were also Western blotted with antibodies to the S epitope or Cdo.