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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).
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