FIGURE 7.

Mutation in either Trp-2 or Ala-80 abolishes cross-linking of hDsc2. A, hDsc2 mutants were incorporated into desmosomes. Confluent cells expressing wild type hDsc2-HA or mutants hDsc2HAW2A or hDsc2HAA80I were double-stained with anti-HA and anti-DP, 11-5F. Co-localization of overexpressed wild type hDsc2 or its mutants with desmoplakin was observed by confocal microscopy. Scale bars, 5 μm. B, Western blot shows that wild type hDsc2 was cross-linked after SEGS treatment but that the mutants, which were expressed in comparable amounts, were not. C, as a control, an identical copy of the blot shown in B was stained with anti-hDsc2 antibody, showing that the endogenous Dsc2 was cross-linked in the presence of both mutants.