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Figure 7. β-Catenin not phosphorylated at NH2-terminal GSK-3β sites binds to cadherin. (A) Cytosolic fraction from HEK293 cells ± Wnt3a was affinity precipitated with cad-GST and TCF-GST, and blotted with pAbs to β-catenin (top blot) or NH2-terminal unphosphorylated–β-catenin (amino acids 27–37, bottom blot). (B) NH2-terminal unphospho–β-catenin localizes to sites of cell–cell contact in Wnt-expressing cells. Rat1 fibroblasts ± Wnt were fixed and processed for immunofluorescence using standard protocols. Images were captured with the Axioplan 2 microscope and AxioVision2.0 software (Carl Zeiss MicroImaging, Inc.). Note that membrane staining of the unphospho-β-catenin (Cy3) is more readily detected under methanol, rather than PFA fixation conditions, perhaps accounting for the apparent differences observed between our study and Staal et al. (2002). Image published in: Gottardi CJ and Gumbiner BM (2004) Copyright © 2004, The Rockefeller University Press. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license Larger Image Printer Friendly View

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