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Fig. 5. Cortactin is important for N-cadherin-mediated intercellular adhesion and surface expression. (A) Cortactin gene silenced cells show a twofold reduction of intercellular adhesion compared with GFP controls (P<0.05). Data are from n=4 replicate samples, means±s.e.m. Immunofluorescence analysis shows lack of cortactin staining and alteration of actin filament network in cortactin knockdown cells. Bar, 20 µm. Immunoblotting shows the extent of cortactin knockdown by RNAi. Samples V, G and SR represent vehicle controls, GFP-RNAi transfected cells, and cortactin RNAi-transfected cells, respectively. Individual samples are corrected for total protein concentration and co-blotted for B-actin. (B) N-cadherin surface labeling of unpermeabilized cells reveals ~25% greater labeling in cortactin RNAi compared with GFP RNAi-treated samples (P<0.05). Permeabilized samples show ~85% reduction of cortactin expression in cortactin RNAi-treated vs. GFP RNAi-treated samples (P<0.005). (C) No effect noted of cortactin siRNA treatment on fibroblast spreading on fibronectin when compared with GFP siRNA-treated controls. (D) No effect noted of cortactin siRNA treatment on strength of cell-substratum adhesion when compared with GFP siRNA-treated controls.





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