FIG. 4.

β-Catenin–LEF-1–DNA complex formation in plakoglobin-transfected cells. (A) Schematic representation of N- and C-terminally VSV-tagged catenin constructs. (B) Catenin–LEF-1–DNA complex formation was analyzed with nuclear extracts from 293-T cells transfected with LEF-1 and VSV-plakoglobin, or VSV–β-catenin. Gel EMSAs were performed with 6 μg of nuclear extract from 293-T cells transfected with control vector (lane 1) or with LEF-1 and the indicated catenin mutants. Nuclear extracts were incubated with a ³²P-labeled DNA probe containing the LEF-1 binding site, in the absence or presence of the indicated antibodies. DNA-protein complexes were analyzed as described in the legend to Fig. 2. N. S., nonspecific. (C) The nuclear extracts used in panel B were analyzed, by immunoblotting, for levels of catenins (with anti-VSV antibody [upper panel]) and LEF-1 (with anti-HA antibody [lower panel]).