Figure 1.
Lateral and apical AJs in A431 and DLD1 cells recruit different combinations of p120 and pkp4. (a) Projections of all x–y optical slices of A431 cells triple stained for E-cadherin (Ecad), p120, and pkp4. The merged image shows p120 (green) and pkp4 (red). Bar, 20 μm. The region outlined by the blue dashed box is enlarged at the right. Arrows highlight pkp4-enriched clusters, which either completely lack p120 (bottom arrow) or be embedded within p120-rich apical AJs (top arrow). Bar, 5 μm. (b) The region marked by the white dashed box in a is shown in three staining combinations: p120 (red)/Ecad (green), pkp4 (red)/Ecad (green), and pkp4 (red)/p120 (green). Note that p120 and E-cadherin are present in nearly all AJs, whereas pkp4 is predominantly enriched in the submicron size dot-like AJs. Scale bar, 20 μm. Apical (ap) and basal (bl) ends of the lateral membranes are indicated. Corresponding scatterplots of relative pixel intensities are shown on the right; Pearson’s r values appear in the upper right of each plot. Note that p120/Ecad pixels cluster within a region of strong positive correlation (shaded zone between red lines), whereas pkp4/Ecad and pkp4/p120 show weaker correlations. (c) Projections of optical z-slices spanning the apical, middle, and basal regions of the cell shown in b. The apical projection spans the top 0.9 μm, the middle projection the next −1.5 μm, and the basal projection—the bottom 0.6 μm of the cell. Bar, 20 μm. Note that p120 is concentrated at apical AJs (white arrows) and basal AJs (arrowhead), whereas pkp4 is enriched in dot-like AJs in the middle portion of the contact (one example indicated by a blue arrow). (d) Maximum-intensity projection of all x–y slices of DLD1 cells stained for p120 (green) and pkp4 (red). Only the merged image is shown. Bar, 20 μm. A zoom of the dashed box is shown at bottom left (bar, 10 μm), and a further magnified view of its dashed region is shown at the right (bar, 5 μm). (e) Scatterplots of p120/pkp4 relative pixel intensities for apical (green) and lateral (red) optical slices of the cell shown in d. Pearson’s r values for each pixel population are indicated in the upper right.

Lateral and apical AJs in A431 and DLD1 cells recruit different combinations of p120 and pkp4. (a) Projections of all x–y optical slices of A431 cells triple stained for E-cadherin (Ecad), p120, and pkp4. The merged image shows p120 (green) and pkp4 (red). Bar, 20 μm. The region outlined by the blue dashed box is enlarged at the right. Arrows highlight pkp4-enriched clusters, which either completely lack p120 (bottom arrow) or be embedded within p120-rich apical AJs (top arrow). Bar, 5 μm. (b) The region marked by the white dashed box in a is shown in three staining combinations: p120 (red)/Ecad (green), pkp4 (red)/Ecad (green), and pkp4 (red)/p120 (green). Note that p120 and E-cadherin are present in nearly all AJs, whereas pkp4 is predominantly enriched in the submicron size dot-like AJs. Scale bar, 20 μm. Apical (ap) and basal (bl) ends of the lateral membranes are indicated. Corresponding scatterplots of relative pixel intensities are shown on the right; Pearson’s r values appear in the upper right of each plot. Note that p120/Ecad pixels cluster within a region of strong positive correlation (shaded zone between red lines), whereas pkp4/Ecad and pkp4/p120 show weaker correlations. (c) Projections of optical z-slices spanning the apical, middle, and basal regions of the cell shown in b. The apical projection spans the top 0.9 μm, the middle projection the next −1.5 μm, and the basal projection—the bottom 0.6 μm of the cell. Bar, 20 μm. Note that p120 is concentrated at apical AJs (white arrows) and basal AJs (arrowhead), whereas pkp4 is enriched in dot-like AJs in the middle portion of the contact (one example indicated by a blue arrow). (d) Maximum-intensity projection of all x–y slices of DLD1 cells stained for p120 (green) and pkp4 (red). Only the merged image is shown. Bar, 20 μm. A zoom of the dashed box is shown at bottom left (bar, 10 μm), and a further magnified view of its dashed region is shown at the right (bar, 5 μm). (e) Scatterplots of p120/pkp4 relative pixel intensities for apical (green) and lateral (red) optical slices of the cell shown in d. Pearson’s r values for each pixel population are indicated in the upper right.

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