Figure 1.

Preferred association between Dvl2 and WWP2. (a) Western blots of coIPs from HEK293ET cells co-transfected with wt or PPxY mutant Flag-Dvl2 and different members of the NEDD4 family (tagged with haemagglutinin (HA)), showing PPxY-dependent association of Dvl2 with these E3 ligases except for WWP2 whose high affinity for Dvl2 only partially depends on its PPxY element. (b) Single confocal sections of representative HeLa cells co-transfected with Dvl2 and the indicated E3 ligases (bottom three rows) or the ligases alone (upper row), and co-stained with α Flag (red) and α HA (green), showing efficient recruitment of cytoplasmic WWP2 into the punctate Dvl2 signalosomes (as judged by the almost complete absence of diffuse HA staining, asterisk), which is blocked by the PPxY mutation (PYm); blue, DAPI stain; scale bars, 10 µM. (c,d) Quantification of IF data shown in (b); 100 cells were scored in each case and assigned to one of three classes (punctate, e.g. WWP2; punctate and diffuse, e.g. NEDD4; diffuse, e.g. NEDD4L).