Fig. 2: MAGI proteins regulate apical contractility and Par-3 localization to AJC. | Communications Biology

Fig. 2: MAGI proteins regulate apical contractility and Par-3 localization to AJC.

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

Fig. 2

a Representative immunofluorescence images of a co-culture of WT and ZO-1,-2 DKO cells stained for either MAGI-1 (magenta, upper panel) or MAGI-3 (magenta, lower panel) with ZO-1 (green). Scale bar, 10 μm. b Pseudocolor representations of apical areas in MAGI-1,-3 DKO cells treated with either DMSO or 10 μM Y-27632 for 5 h. c Violin plot depiction of apical areas in MAGI-1,-3 DKO cells. Data were collated from six measurements obtained over three independent experiments. Lines represent the median and the upper and lower quartiles. P value from unpaired t test is shown. n = 164 (DMSO) and 147 (Y-27632). d Frequency distributions of apical areas in WT and MAGI-1,-3 DKO cells. Data were as in c. Points are means and error bars are SDs. e P values of D’Agostino & Pearson test for normal distribution were computed from the data presented in d. Medians are notated and shown graphically as lines. f Representative immunofluorescence images of WT and MAGI-1,-3 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. g Cross-junctional line scans of ROCK1 immunofluorescence from images corresponding to f. 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. h Representative immunofluorescence images of a co-culture of WT and MAGI-1,-3 DKO cells stained for Par-3 (magenta) and MAGI-3 (green). Orthogonal views of the cross-section indicated by the white dotted line are shown to the right of each image. Scale bar, 10 μm. i Cross-junctional line scans of Par-3 immunofluorescence in WT and MAGI-1,-3 DKO cells. Cells were co-stained with 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. j Quantification of Par-3 mean fluorescence intensities at AJC in WT and MAGI-1,-3 DKO cells from images common with i. P value from unpaired t test is shown. k Quantification of Par-3 coverage relative to total AJC area based on data obtained with j. P value from unpaired t test is shown. Source data are available in Supplementary Data 1.

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