Figure S1.
Pkp4 is enriched at lateral AJs. (a) Ratio of pkp4 to E-cadherin fluorescence intensities (pkp4/Ecad index) assessed for lateral, apical, and basal AJs (20 junctions each) across four cell–cell contact regions from two independent images. Pkp4 levels are significantly higher in lateral AJs compared with apical or basal AJs. Data are presented as mean ± SD. Statistical significance was determined using a two-tailed Student’s t test, ****P <0.0001. (b) Colocalization color maps depicting colocalization between E-cadherin-p120, E-cadherin-pkp4, and p120-pkp4. The −1 to +1 heat map displays the nMDP, which represents correlation between intensities of corresponding pixels: negative correlation (-1-0) are shown in blue-green colors, whereas positive correlations (0 to +1) are represented by yellow-red colors. The index of correlation (Icorr) for each protein pair is shown in the upper right corner. Bar, 20 μm. Consistent with Pearson’s analyzes (Fig. 1), this approach reveals the strongest correlation between E-cadherin and p120, and the weakest between p120 and pkp4. It also shows that apical AJs contain isolated pkp4-enriched clusters (one example is indicated by an arrow in the zoomed images at the bottom panel). Bar, 10 μm. (c) Maximum-intensity projections of A431 cells stained for pkp4 (green) and for desmosomal protein dsg2 (dsg2, red). Only the merged image is shown at low magnification. Bar, 20 μm. Zoomed regions (dashed boxes) are shown at the bottom panel and in the inset for each channel separately. Bar, 10 μm. Note that DSMs do not colocalize with pkp4-rich AJs, although they are frequently adjacent. (d) Scatterplot of red versus green relative pixel intensities for the image in panel c. The PCC (r) is indicated in the upper right corner. Note the nearly complete absence of positive correlation. (f) MSD measured from a region containing ∼16 lateral AJs from Video 1 (black trace). Note that the observed MSD deviates from a linear trend (red line) suggesting the active, non-Brownian movement of the junctions.

Pkp4 is enriched at lateral AJs. (a) Ratio of pkp4 to E-cadherin fluorescence intensities (pkp4/Ecad index) assessed for lateral, apical, and basal AJs (20 junctions each) across four cell–cell contact regions from two independent images. Pkp4 levels are significantly higher in lateral AJs compared with apical or basal AJs. Data are presented as mean ± SD. Statistical significance was determined using a two-tailed Student’s t test, ****P <0.0001. (b) Colocalization color maps depicting colocalization between E-cadherin-p120, E-cadherin-pkp4, and p120-pkp4. The −1 to +1 heat map displays the nMDP, which represents correlation between intensities of corresponding pixels: negative correlation (-1-0) are shown in blue-green colors, whereas positive correlations (0 to +1) are represented by yellow-red colors. The index of correlation (Icorr) for each protein pair is shown in the upper right corner. Bar, 20 μm. Consistent with Pearson’s analyzes (Fig. 1), this approach reveals the strongest correlation between E-cadherin and p120, and the weakest between p120 and pkp4. It also shows that apical AJs contain isolated pkp4-enriched clusters (one example is indicated by an arrow in the zoomed images at the bottom panel). Bar, 10 μm. (c) Maximum-intensity projections of A431 cells stained for pkp4 (green) and for desmosomal protein dsg2 (dsg2, red). Only the merged image is shown at low magnification. Bar, 20 μm. Zoomed regions (dashed boxes) are shown at the bottom panel and in the inset for each channel separately. Bar, 10 μm. Note that DSMs do not colocalize with pkp4-rich AJs, although they are frequently adjacent. (d) Scatterplot of red versus green relative pixel intensities for the image in panel c. The PCC (r) is indicated in the upper right corner. Note the nearly complete absence of positive correlation. (f) MSD measured from a region containing ∼16 lateral AJs from Video 1 (black trace). Note that the observed MSD deviates from a linear trend (red line) suggesting the active, non-Brownian movement of the junctions.

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