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Fig. 5. Calcium-induced migfilin localization to adherens junction. (A-D) MCF7 mammary epithelial cells were subject to calcium-chelation assay as described in Materials and Methods. The cells were fixed at 15 minutes (A and B) and 30 minutes (C and D) after switching to calcium-containing medium, and double stained with a rabbit anti-ß-catenin antibody (A and C) and the mouse monoclonal anti-migfilin antibody (B and D). (E,F) MCF7 mammary epithelial cells were transiently transfected with an expression vector encoding GFP-migfilin. The cells were subject to calcium-chelation assay in the presence of a function-blocking mouse monoclonal anti-E-cadherin antibody (SHE78-7) (G-J) or a non-specific mouse IgG as a control (E and F) as described in Materials and Methods. The cells were fixed at 60 minutes after switching to calcium-containing medium, and stained with a rabbit anti-ß-catenin antibody (E, G and I) and a Rhodamine Red^TM-conjugated anti-rabbit IgG antibody. ß-Catenin (E, G and I) and GFP-migfilin (F, H and J) at the cell-cell (E-H) or cell-ECM (I and J) adhesions were detected using a Leica DM R fluorescence microscope. Bars, 5 µm.

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