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Figure 2


Fig. 2. Wnt-dependent relocation of Dvl2-GFP and endogenous Dvl2 puncta to the plasma membrane. (A,B) Fixed 293T cells, after transfection with Dvl2-GFP (50 ng), Fz8 (100 ng), LRP6 (100 ng), GSK3beta (100 ng) and HA-axin (100 ng), (A) before or (B) after 2 hours of exposure to recombinant Wnt3a, showing Wnt-dependent recruitment of Dvl2-GFP to the plasma membrane. (C-F) 293T cells, (C-E) before or (F) after exposure to Wnt3a as in B, fixed and stained with affinity-purified anti-Dvl2 antiserum, revealing endogenous cytoplasmic Dvl2 puncta (C) that are recruited to the plasma membrane (F, arrows) in a Wnt-dependent way (insert in C, high-magnification view of cytoplasmic puncta of endogenous Dvl2); the specificity of the signals is indicated by the near-complete elimination of the staining after (D) siRNA treatment against Dvl2, or (E) prior depletion of the antiserum with 100x molar excess of recombinant GST-DIX. Scale bar, 15 µm.