Fig. 7. Wnt signalling does not influence ß-catenin nucleo-cytoplasmic shuttling. (A) Immunofluorescence staining of endogenous ß-catenin (upper panels) in HEK293T cells in the absence or presence of mRFP-tagged Wnt-3a or dominant active LRP6 (LRP6da). Lower panels depict the fluorescence of the RFP-tagged activators in the same microscopic field as in the upper panels. Bar, 10 µm. (B) Western blot of hypotonic lysates of HEK293T cells transfected with mYFP-ß-catenin and the indicated activators, stained for ß-catenin (top) and ß-actin (bottom). (C) TOPglow reporter assay with mYFP-ß-catenin and indicated interactors. Fold changes were determined by calculating ratios of relative luciferase activities of TOP- to FOP samples and normalizing to the mYFP control. Error bars indicate standard deviations. (D) Compartment bleach analysis of mYFP-ß-catenin in the absence or presence of dominant active LRP6 or Wnt3a. The cartoon designates compartment bleaches.