Fig. 3: ASPP2 is recruited to AJC by MAGIs, where it regulates apical contractility and Par-3 localization to AJC. | Communications Biology

Fig. 3: ASPP2 is recruited to AJC by MAGIs, where it regulates apical contractility and Par-3 localization to AJC.

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

Fig. 3

a Representative immunofluorescence images of WT and MAGI-1,-3 DKO cells stained for ASPP2 (magenta) and MAGI-3 (green). Scale bar, 10 μm. b Cross-junctional line scans from immunofluorescence images of WT and MAGI-1,-3 DKO cells stained for ASPP2 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. c Immunoblot of FLAG immunoprecipitation showing interaction between HA-MAGI-3 and FLAG-ASPP2, either with or without GFP-RASSF10. Data is representative of three independent experiments. Uncropped immunoblots are shown in Supplementary Fig. S7. d Quantification of c. The amount of HA-MAGI-3 coprecipitating with FLAG-ASPP2 in the presence of GFP-RASSF10 was normalized to that of HA-MAGI-3 coprecipitation in the presence of control GFP. P value from unpaired t test is shown. e Pseudocolor representations of apical areas in ASPP1,2 DKO cells treated with either DMSO or 10 μM Y-27632 for 5 h. f Violin plot depiction of apical areas in WT and ASPP1,2 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 = 168 (DMSO) and 131 (Y-27632). g Frequency distributions of apical areas in WT and ASPP1,2 DKO cells. Data were as in f. Points are means and error bars are SDs. h P values of D’Agostino & Pearson test for normal distribution were computed from the data presented in g. Medians are notated and shown graphically as lines. i Representative immunofluorescence images of a co-culture of ASPP1,2 DKO and GFP-ASPP2-WTres cells stained for Par-3 (magenta). Scale bar, 10 μm. j Cross junctional line scans of Par-3 immunofluorescence in ASPP1,2 DKO, and GFP-ASPP2-WTres cells. ASPP1,2 DKO cells were co-stained with activated α-catenin. Shaded area represents AJC as defined by either the activated α-catenin or GFP peak. Individual data from 20 independent line scans are shown with the means depicted by solid lines. k Quantification of Par-3 mean fluorescence intensities at AJC in WT and ASPP1,2 DKO cells from images common with j. P value from unpaired t test is shown. l Quantification of Par-3 coverage relative to total AJC area based on data obtained with k. P value from unpaired t test is shown. Source data are available in Supplementary Data 1.

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