FIG. 7.

Immunocytochemistry shows that the primary and NCI-N87 cells show similar patterns of E-cadherin and actin staining. All three cell preparations were grown on APES-coated coverslips for 48 h prior to fixation with 4% paraformaldehyde and permeabilization with 0.1% Triton X-100. All cells were immunostained with a monoclonal anti-E-cadherin antibody for 18 h at 4°C before being incubated with phalloidin-AlexaFluor 594 for 1 h at room temperature. At the basolateral membrane E-cadherin (A, primary; E, NCI-N87) is reduced and actin stress fibers predominate (B, primary; F, NCI-N87). From 1 μm from the substrate to the top of the cells both showed strong E-cadherin (C, primary; G, NCI-N87) and cortical actin (D, primary; H, NCI-N87) staining at the plasma membrane. AGS cells show a punctate pattern of E-cadherin staining that does not localize to adherens junctions (I). AGS cells have a uniform pattern of filamentous actin staining with a small number of stress fibers (arrows) and cortical actin (arrowheads) (J). Scale bar = 10 μm. Images are representative of three individual experiments.