Figure 1

Characterization of vinculin knock-down cells. (A) Western blot of detergent soluble and insoluble pool of vinculin in wild-type (WT) and vinculin knockdown (KD) cells. (B) Quantification of total and detergent insoluble pool of vinculin in wild-type and knockdown cells. The fraction of detergent insoluble (pellet) vinculin pool increased in both vinculin knockdown cells. (C) Immuno-fluorescence analysis of vinculin in wild-type and knockdown cells. Although vinculin is depleted from the cytoplasm of knockdown cells, the residual vinculin is present at focal adhesions (top panel) and cell-cell contacts (bottom panel). Images were taken at basal (top panel) and apical (bottom panel) levels. The same exposure and laser power were used to acquire and generate the images. Scale bar 10 μm. (D) Quantification of vinculin at focal adhesions in wild-type and knockdown cells. Immuno-fluorescence intensities and analysis of vinculin at focal adhesions are presented as mean ± standard error of the mean and are compared with control (*P < 0.05). (E) Quantification of vinculin at cell-cell contacts in wild-type and knockdown cells. Immuno-fluorescence intensities and analysis of vinculin at cell-cell contacts are presented as mean ± standard error of the mean and are compared with control (***P < 0.001).