Fig. 1: Loss of ZO proteins dysregulates ROCK-dependent contractility to alter apical morphology. | Communications Biology

Fig. 1: Loss of ZO proteins dysregulates ROCK-dependent contractility to alter apical morphology.

From: MAGIs regulate aPKC to enable balanced distribution of intercellular tension for epithelial sheet homeostasis

Fig. 1

a Representative immunofluorescence images of WT and ZO-1,-2 DKO cells stained for activated α-catenin. Scale bar, 20 μm. b Representative immunofluorescence images of a co-culture of WT and ZO-1,-2 DKO cells stained for phosphorylated MLC (pMLC, magenta) and ZO-1 (green). Scale bar, 10 μm. c Cross-junctional line scans (pMLC) from immunofluorescence images of WT and ZO-1,-2 DKO cells stained for pMLC and activated α-catenin. Shaded area represents AJC as defined by the activated α-catenin peak. Individual data from 20 independent line scans are shown with the means depicted by solid lines. d Pseudocolor representations of apical areas in WT and ZO-1,-2 DKO cells. ZO-1,-2 DKO cells were treated with either DMSO or 10 μM Y-27632 for 5 h. Cells were stained for activated α-catenin and processed for area measurement as detailed in Supplementary Fig. S1 and Methods. e Violin plot depiction of apical areas in WT and ZO-1,-2 DKO cells. Areas were collated from six measurements obtained over three independent experiments. Lines represent the median and the upper and lower quartiles. P values from Tukey’s post-hoc test with one-way ANOVA are shown. n = 228 (WT), 219 (ZO-1,-2 DKO DMSO) and 191 (ZO-1,-2 DKO Y27632). f Frequency distributions of apical areas in WT and ZO-1,-2 DKO cells. Data were as in e. Points are means and error bars are SDs. g P values of D’Agostino & Pearson test for normal distribution were computed from the data presented in f. Medians are notated and shown graphically as lines. h Representative immunofluorescence images of WT and ZO-1,-2 DKO cells stained for ROCK1 (magenta) and activated α-catenin (green). Orthogonal views of the cross-section indicated by the yellow dotted line are shown to the right. Scale bar, 10 μm. i Cross-junctional line scans of ROCK1 immunofluorescence from images corresponding to j. Shaded area represents AJC as defined by the activated α-catenin peak. Individual data from 20 independent line scans are shown with the means depicted by solid lines. j Representative immunofluorescence images of a co-culture of WT and ZO-1,-2 DKO cells stained for Par-3 (magenta) and ZO-1 (green). Scale bar, 10 μm. k Quantification of Par-3 junctional coverage relative to total AJC area in WT and ZO-1,-2 DKO cells. Cells were co-stained for Par-3 and activated α-catenin, which was used to designate AJC. P value from unpaired t test is shown. l Quantification of Par-3 mean fluorescence intensities at AJC based on the same data as j. P value from unpaired t test is shown. Source data are available in Supplementary Data 1.

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