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Figure 6


Fig. 6. Recruitment of CKI{epsilon} into Dvl2 puncta. (A) SuperTOP assays in transfected 293 cells as in Fig. 5A; limiting amounts of HA-CKI{epsilon} (100 ng) were co-expressed with full-length FLAG-Dvl2 (200 ng) or truncations (500 ng), as indicated. (B-E) HeLa cells, cotransfected with HA-CKI{epsilon} (green) and FLAG-Dvl2 or truncations (red) as indicated in panels, fixed and stained as in Fig. 5. Note that Dvl299 does not affect the HA-CKI{epsilon} staining, which remains diffuse cytoplasmic (D), indistinguishable from the staining of HA-CKI{epsilon} expressed by itself, or co-expressed with {Delta}DIX (not shown). (F) SuperTOP assays in 293T cells transfected with FLAG-Dvl2, after RNAi-mediated depletion of endogenous CKI{epsilon} (to <50% of normal levels, as show by western blotting, G), revealing that the signalling activity of Dvl2 depends on CKI{epsilon}. Scale bar, 15 µm.





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