Fig. 9. The phenotype induced by p120ctn isoforms and deletion constructs transiently transfected into HaCaT cells. 16 hours after the transfection, a serum-free medium containing 1 mM Ca²⁺ was added and cells were incubated for 8 hours before preparing for the immunofluorescence staining. The expression of the FLAG-tagged polypeptides was demonstrated with FLAG-tag-specific rabbit antibody (FITC, green signal), endogenous E-cadherin is shown as red signal (Texas Red), and nuclei were visualized with DAPI staining (blue signal). Isoforms 1A (A,B,E,F), 2A (I,J,L,M) and 3A (O,P,S,T), as well as the deletion construct 2ANS (K,N), colocalized with E-cadherin at the cell-cell junctions, but the overexpression of each transgene resulted in the branching phenotype and the simultaneous disappearance of the E-cadherin staining both from the cell-cell adhesion borders as well as from the cytoplasmic pool (arrowheads in K and N). The isoforms 4A (R, V) and 1AB (C,G), as well as the deletion construct 3AAK (Q,U), displayed cytoplasmic localization and did not interfere with the endogenous E-cadherin signal. The C-terminally truncated isoform 1, 1X, (D,H) localized to the nucleus but did not affect the E-cadherin signal. Bar, 50 µm. The bar in (A) applies to panels E,I,K,L,N,O,S, and the bar in (B) applies to panels C,D,F,G,H,J,M,P,Q,R,T,U,V.

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