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Fig. 5. Wg signaling leads to redistribution of Arm and to loss of E-cadherin from the cell surface. cl8Wg^ts/mEcad cells incubated for 6 hours at the restrictive temperature (25°C; A,C) or the permissive temperature (16°C; B,D) in the presence of Cu²⁺ were triple stained with antibodies against Arm (green), E-cadherin (red) and Wg (blue). (A) At 25°C, when Wg^ts protein is nonfunctional, mutant Wg exhibited a diffuse, perinuclear distribution. Arm and E-cadherin colocalized at cell-cell contacts and in intracellular dots, presumably vesicles. (B) At 16°C, Wg^ts protein is active and developed a punctate staining pattern. Arm was no longer localized to cell-cell contacts but was enriched in the cytoplasm and at the tips of cellular processes that appeared to be in close contact with the substratum. E-cadherin staining was strongly reduced at the cell surface and was most prominent in the perinuclear region. Little colocalization of Arm and E-cadherin was visible. (C,D) Superimposition of Arm staining (top) with phase-contrast image (bottom) of cells. (C) At the nonpermissive temperature cells formed tight contacts with each other and did not extend membrane processes. (D) Cells at the permissive temperature had less developed cell-cell contacts and extended many processes that contained high amounts of Arm. Bar, 10 µm.

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