Figure 5. Confocal microscopy to investigate the localization of F-actin, E-cadherin and vinculin in the cellular islands.

MDCK cell islands were imaged using confocal microscopy to examine the location of these proteins through the thickness. On the basis of nuclear location, the basal section (0.1~0.4 μm) was regarded as the region of focal adhesions (FAs), and the mid-apical section (0.9~1.2 μm) was regarded as the region of adherens junctions (AJs). As in Fig. 4, white arrows indicate co-localization of each protein and yellow arrows indicate dissociation. Representative regions used to examine co-localization (white boxes) are enlarged on the upper right panel (white; locations of co-localization) and presented in a scatter plot with a correlation coefficient line on the lower right panel. (a) E-cadherin was present only on AJ region because it was not expressed in FA region. (Red; E-cadherin, green; F-actin, and blue; Hoechst) (b) Localization of vinculin and F-actin showed differential expression depending on the z-position. F-actin and vinculin show lower expression in the AJ region in HGF sample compared to the control sample. (Red; vinculin, green; F-actin, and blue; Hoechst).