Fig. 9. Rescue of D743R cells by expression of wild-type Fer or Y654F ß-catenin. (A) D743R cells were transfected with wild-type Fer cDNA and stable cell lines were selected. Equivalent amounts of protein from cell lysates were analysed by immunoblotting with anti-Fer antibody. `D743R+WTFer' indicates lysates of D743R cells transfected with wild-type Fer cDNA. (B) WT, D743R and D743R cells transfected with wild-type Fer cDNA (D743R + WT) were lysed in neutral detergent, immunoprecipitated with anti-pan-cadherin antibody and analysed by western blot with anti-ß-catenin and anti-PTP1B antibodies. (C) Cells grown on multiple-well slides were fixed, permeabilized and stained with Alexa-568/phalloidin and anti-cadherin or anti-ß-catenin antibody followed by Alexa-488-labeled secondary antibody. Notice that cadherin and ß-catenin are again localized at cell boundaries in D743R cells expressing wild-type Fer. Scale bar, 10 µm. (D) D743R cells were incubated with GST/Y654F-ß-catenin, GST/wild-type-ß-catenin, or GST/Y142F-ß-catenin in the presence of BioPorter, the cells were lysed, immunoprecipitated with anti-GST antibody and immunoblotted with anti-pan-cadherin antibody. The immunoblots were stripped and reprobed with anti-GST antibody. (E) D743R cells expressing GST/Y654F-ß-catenin were visualized under phase contrast and photographed using an Axiovert 25 microscope and Axiovision (Zeiss) after 3 hours at 37°C in 30 mm dishes rotated at 80 rpm.