Figure 1.

(A) Differences in the degree of cell aggregation seen in sub-confluent cultures of cells expressing the Naxos mutant compared with cells expressing wildtype (wt) or the insS mutant form of plakoglobin. The Naxos cells are more dispersed (cluster less) than either the wildtype or insS mutant cells. Magnification is 4x; image digitally magnified 2x, scale bars = 200 μm. (B) Western blot showing roughly equal amounts of wildtype and mutant plakoglobin expressed in transfected cell lines. The Naxos protein migrates with a lower molecular weight, consistent with the premature termination. Numbers on the left represent molecular weights of a standard ladder. (C) Western blot using a C-terminal-region antibody against plakoglobin showing that the Naxos plakoglobin band is absent, consistent with C-terminal truncation, but also showing a faint band in the Naxos cell lane which presumably represents endogenous (wildtype) plakoglobin. (D) Confocal images of cells stained with an antibody recognizing a C-terminal epitope of plakoglobin. There is abundant junctional localization of plakoglobin in the wildtype and insS mutant cells but not in the Naxos cells. Scale bar = 10 μm. (E) Confocal images of cells stained with an antibody recognizing an N-terminal epitope of plakoglobin. Again, there is abundant junctional localization of plakoglobin in the wildtype and insS mutant cells whereas in the Naxos cells there is nuclear localization. Scale bar = 10 μm.