Fig. 3. Point mutations decrease the interaction between Dsh and HFz5. (A) Recruitment of Dsh to the plasma membrane by wild-type and mutant human FZ5. Myc-tagged Dsh and GFP-tagged human FZ5 were co-expressed in COS cells. Cells were stained by anti-Myc 9E10 monoclonal antibodies and Texas Red-conjugated goat anti-mouse secondary antibodies, and examined by fluorescence microscopy. (B) Physical interaction between Dsh and wild-type or mutant human FZ5. Myc-tagged Dsh and GFP-tagged human FZ5 were co-expressed in 293 cells. Cell lysates were immunoprecipitated with anti-Myc antibodies, and immunoprecipitates were fractionated by SDS-PAGE, transferred to a nitrocellulose membrane and immunoblotted with anti-GFP antibodies (upper panel). Expression of Dsh and human FZ5 in total cell lysates was examined by immunoblotting with anti-Myc and anti-GFP antibodies (lower panels).