CAP350 peripheral labelling is dependent on E-cadherin-mediated cell adhesion.

<p>(<b>A</b>) Control cells were treated with 4 mM EGTA for 2 h to disrupt cell-cell contacts, fixed at the indicated times after washout, and stained with anti-E-cadherin and CAP350r (top) or anti-α-catenin and CAP350r (bottom) antibodies. Single labellings are shown. (<b>B</b>) Low-magnification images of MDCKII cells treated with either a control immunoglobulin G (IgG) antibody (Ab) or the E-cadherin blocking Ab (DECMA-1) for 72 h. Treated cells were then fixed and stained for CAP350. The presence of DECMA-1 antibody bound to E-cadherin at the cell surface was visualised with a secondary antibody. (<b>C</b>) Representative images of cells treated as in (B) and double labelled for α-catenin and CAP350. E-cadherin in DECMA-1-treated cells was revealed by incubation with a secondary antibody. Merged images are shown at right. High-magnification images of boxed areas are shown in the panels underneath. (<b>D</b>, <b>E</b>) MDCKII cells processed as in (B) but stained with zonula occludens protein 1 (ZO-1) (<b>D</b>) or α-tubulin antibodies (<b>E</b>). Bars = 10 μm.