Fig. 2.

1 mM 4-PBA results in loss of cell-cell adhesion through diminished cell-cell contacts.

A In the dispase-based dissociation assay the 1 mM 4-PBA-treated cells detached significantly easier resulting in more fragments and thus reflecting disturbed cell-cell contacts [for NHK+, KRT14mut + and KRT5mut+: ***P < .0001, student's t-test].

B Immunofluorescence staining with anti-desmoplakin (red) antibodies to visualize desmosomes, and anti-plakoglobin (green) antibodies to demonstrate adherence junctions, in NHK was performed after calcium switch. Both were reduced and their localization at the cell-cell junctions was lost after treatment with 1 mM 4-PBA. Scale bar: 50 μm.

C Immunoblotting for desmoplakin, E-cadherin and plakoglobin revealed reduced abundance after 1 mM 4-PBA treatment, whereas β-catenin was found enhanced. Bar graphs on the right show expression of these proteins normalized to tubulin as percentage of expression in untreated cells [desmoplakin: P = .2383 for KRT14mut and *P = .0343 for KRT5mut; E-cadherin: P = .0787 for KRT14mut and **P = .0023 for KRT5mut; β-catenin: P = .0675 for KRT14mut and P = .7116 for KRT5mut; plakoglobin: *P = .110 for KRT14mut and *P = .0220 for KRT5mut, all done with paired student's t-test]. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)