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Figure 1


Fig. 1. Cadherin-independent plasma membrane localization of dephospho-β-catenin upon Wnt3A stimulation. (A) E-cadherin-negative cells respond normally to Wnt3a. Luciferase reporter assay in Kep1 (E-cadherin–/–) and Kp6 (E-cadherin+/+) cells using the TCF reporter TOP-TK and the control FOP-TK, normalized for transfection efficiency using pRL-CMV-Renilla. 24 hours after transfection, cells were stimulated overnight with Wnt3a-conditioned or control medium and luciferase activity was measured. (B) (Dephospho) β-catenin levels in Kep1 or Kp6 cells. Cells were induced with Wnt3A protein or control and analyzed 0.5 or 2.5 hours after induction by western blotting using an antibody recognizing all forms of β-catenin (TCAT) or an antibody specific for the N-terminal dephospho form (ABC). Asterisk indicates a crossreacting protein. (C) Subcellular localization of dephospho-β-catenin in E-cadherin positive or negative cells upon Wnt stimulation. Kep1 (E-cadherin–/–) or Kp6 (E-cadherin+/+) cells were induced with Wnt3A protein or control and analyzed 2.5 hours after induction by immunolocalization with an antibody specific for the N-terminal dephospho form (ABC). DAPI was used as a nuclear marker. Note that dephospho-β-catenin levels in E-cadherin–/– cells are much lower than in E-cadherin+/+ cells, requiring unequal confocal settings to be used.





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