Fig. 5
Characterization of E-cadherin/CDCP1-containing protein complexes. a MDA-MB-231 or T47D cell lines stably expressing an E-cadherin-glutathione S-transferase (E-cad-GST) fusion protein were transduced with an adenovirus encoding ΔCDCP1-FLAG or a control adenovirus encoding GFP. Cell extracts were subjected to sequential affinity purification using anti-FLAG agarose followed by glutathione-agarose. b T47D cell extracts were immunoprecipitated with a CDCP1 antibody and the immunoprecipitates were analyzed by immunoblot with the indicated antibodies. c The indicated stable cell lines were transduced with adenoviruses encoding GFP (control) or ΔCDCP1-FLAG and cell extracts were subjected to affinity purification with either anti-FLAG agarose or glutathione agarose. Affinity-purified proteins were analyzed by immunoblot. d HEK 293 cells were transiently transfected with the indicated tagged CDCP1 constructs and treated with the specified DDAs or vehicle. CDCP1 dimerization was monitored by TALON pulldown of His6-tagged proteins followed by purification with anti-FLAG agarose and immunoblot analysis of the sequentially affinity-purified material. e HEK 293 cells were transiently transfected with the indicated plasmid constructs and cell extracts were affinity purified sequentially using TALON resin and anti-FLAG agarose. The affinity-purified material was analyzed by immunoblot