Figure 4

Interrogation of functional roles of Y17 and S407 sites of Dvl3 in formation of macromolecular Dvl3-based punctae in response to stimulation with Wnt3a. Panel A, live cell images of expression of either wild-type Dvl3 or Y17D-Dvl3 or S407A-Dvl3 or S407D-Dvl3 in response to Wnt3a. F9 cells were co-transfected with Rfz1 and either wild-type Dvl3 or Y17D-Dvl3 or S407A-Dvl3 or S407D-Dvl3. One day post transfection, cells were stimulated with Wnt3a and cells images were taken at 0, 15, and 30 min post stimulation with Wnt3a. Panel B, live cell images of cells expressing either wild-type or Dvl3 mutants (Y17D-Dvl3 or S407D-Dvl3 or S407D-Dvl3) in Dvl3-deficient cell: response to Wnt3a. F9 cells stably expressing with M50 and Rfz1 were treated with siRNA targeting Dvl3. On the following day, cells were transfected with either wild-type Dvl3 or one of the Dvl3 mutants. Post 24 hr of transfection, cells were stimulated with Wnt3a and live cell images were captured during Wnt stimulation. Results were displayed at 0, 5, and 10 min time points. The images shown are representative of three or more separate experiments.