DOI: 10.5061/dryad.bvq83bkkb
title: Epithelial polarization by the core planar cell polarity complex is exclusively non-autonomous

description: For cells to polarize collectively along a tissue plane, asymmetrically
 localized planar cell polarity (PCP) complexes must form intercellular
 contacts between neighboring cells. Yet, it is unknown whether asymmetric
 segregation of PCP complexes requires cell-cell contact, or if cell-
 autonomous, antagonistic interactions are sufficient for polarization. To
 test this, we generated mouse chimeras consisting of dual PCP-reporter
 cells mixed with unlabeled cells that cannot form PCP bridges. In the
 absence of intercellular interactions, PCP proteins failed to polarize
 cell autonomously. Rather, PCP-mediated contacts along single cell-cell
 interfaces were sufficient to sort PCP components to opposite sides of the
 junction, independent of junction orientation. Thus, intercellular binding
 of PCP complexes is the critical step that initiates sorting of opposing
 PCP complexes to generate asymmetry.
description: # Epithelial polarization by the core planar cell polarity complex is
 exclusively non-autonomous
 [https://doi.org/10.5061/dryad.bvq83bkkb](https://doi.org/10.5061/dryad.bvq83bkkb) ## Description of the data and file structure This dataset contains all data crucial to the key findings of this study, including original raw images of chimeric embryonic skin (ads5704_RawImages.zip), data analysis (*ads5704_Data.xlsx), and scripts (ads5704_MATLAB scripts.zip). Using embryo aggregation, we generated mouse chimeras to test the hypothesis of whether planar cell polarity (PCP) can arise cell autonomously in the absence of intercelullar PCP interactions. The raw data is uploaded in original format allowing for replication of presented analyses and further analysis or interpretation.  ### Files and variables #### File: ads5704\_Data.xlsx **Description:** Excel file containing all quantification of analyses presented in the publication. Each tab in the excel sheet corresponds with a figure as labeled by figure number. Each analyzed cell, cell pair or group of cells is labeled with a number that remains consistent across excel tabs of the same color.  Fig. 1G (tab 1): all data corresponding to Fig. 1G * *Embryo*: indicates biological replicate * *Single cell number*: cell identifier, corresponds to number of image in RawImages.zip * *Anterior (A) or Posterior (P) cell border*: indicates which border, A or P, is analyzed * *Border mean intensity (Ij)*: mean fluorescence intensity of Fz6-3xGFP, tdTomato-Vangl2 and Celsr1 along individual cell borders * *Cell mean intensity (Ic)*: mean fluorescence intensity of Fz6-3xGFP, tdTomato-Vangl2 and Celsr1 of the cell * *Junction enrichment (Ij/Ic)*: ratio of border mean intensity/cell mean intensity  Fig. 1H (tab 2): all data corresponding to Fig. 1H * *Embryo*: indicates biological replicate * *Single cell number*: cell identifier, corresponds to number of image in RawImages.zip * *Pearson's coefficient*: calculated pearson's correlation coefficient between Fz6-3xGFP and tdtomato-Vangl2; Fz6-3xGFP and Celsr1; and tdtomato-Vangl2 and Celsr1 along cell border of single cell in *Fz6-3xGFP; tdTomato-Vangl2 : WT* chimeric skin Fig. 2E (tab 3): all data corresponding to Fig. 2E * *Embryo*: indicates biological replicate * *Single cell number*: cell identifier, corresponds to number of image in RawImages.zip * *Pearson's coefficient*: calculated pearson's correlation coefficient between Fz6-3xGFP and tdtomato-Vangl2; Fz6-3xGFP and Celsr1; and tdtomato-Vangl2 and Celsr1 along cell border of single cell in *Fz6-3xGFP; tdTomato-Vangl2 : Celsr1-/-* chimeric skin Fig. 3D (tab 4): all data corresponding to WT:WT junctions in Fig. 3D * *Embryo*: indicates biological replicate * *Cell Pair Number*: cell identifier, corresponds to number of image in RawImages.zip * *Angle of junction*: angle of analyzed junction relative to the AP axis * *Border mean intensity (Ij)*: mean fluorescence intensity of Fz6-3xGFP, tdTomato-Vangl2 and Celsr1 along WT:WT cell junctions between dual-reporter cell pairs in in *Fz6-3xGFP; tdTomato-Vangl2 : Celsr1-/-* chimeric skin * *Cell pair mean intensity (Ic)*: mean fluorescence intensity of Fz6-3xGFP, tdTomato-Vangl2 and Celsr1 of the cell pair * *Junction enrichment (Ij/Ic)*: ratio of WT:WT border mean intensity/cell pair mean intensity  Fig. 3D (tab 5): all data corresponding to WT:mutant junctions in Fig. 3D * *Embryo*: indicates biological replicate * *Cell Pair Number*: cell identifier, corresponds to number of image in RawImages.zip * *Angle of junction*: angle of analyzed junction relative to the AP axis * *Border mean intensity (Ij)*: mean fluorescence intensity of Fz6-3xGFP, tdTomato-Vangl2 and Celsr1 along WT:mutant cell junctions between dual-reporter cells and surrounding mutant cells in in *Fz6-3xGFP; tdTomato-Vangl2 : Celsr1-/-* chimeric skin * *Cell pair mean intensity (Ic)*: mean fluorescence intensity of Fz6-3xGFP, tdTomato-Vangl2 and Celsr1 of the cell pair * *Junction enrichment (Ij/Ic)*: ratio of WT:mutant border mean intensity/cell pair mean intensity  Fig. 3E (tab 6): all data corresponding to Fig. 3E * *Embryo*: indicates biological replicate * *Cell Pair Number*: cell identifier, corresponds to number of image in RawImages.zip * *Junction*: indicates analysis of either WT:WT junctions between cell pairs or WT:mutant junctions  * *Pearson's coefficient*: calculated pearson's correlation coefficient between Fz6-3xGFP and tdtomato-Vangl2; Fz6-3xGFP and Celsr1; and tdtomato-Vangl2 and Celsr1 along WT:WT or WT:mutant cell borders in *Fz6-3xGFP; tdTomato-Vangl2 : Celsr1-/-* chimeric skin Fig. 4D (tab 7): all data corresponding to Fig. 4D * *Embryo*: indicates biological replicate * *WT:WT junction number*: junction identifier, corresponds to number of image in RawImages.zip * *Angle of junction*: angle of analyzed junction relative to the AP axis, all junctions are "vertical" with angles between 45-90 * *Number of puncta pairs*: counted number of puncta pairs in either F:V, V:F or ambiguous orientations per WT junction in large area of WT cells in *Fz6-3xGFP; tdTomato-Vangl2 : Celsr1-/-* chimeric skin * *Percentage of puncta pair orientations*: percentages calculated from the counted puncta pairs in the previous columns Fig. 4M,N (tab 8): all data corresponding to Fig. 4D * *Embryo*: indicates biological replicate * *WT:WT junction number*: junction identifier, corresponds to number of image in RawImages.zip * *Angle of junction*: angle of analyzed junction relative to the AP axis. All junctions with angles between 45-90 are considered vertical, while all junctions with angles between 0-45 are considered horizontal. * *Number of puncta pairs*: counted number of puncta pairs in either F:V, V:F or ambiguous orientations per WT:WT junction between dual-reporter cell pairs in *Fz6-3xGFP; tdTomato-Vangl2 : Celsr1-/-* chimeric skin * *Percentage of puncta pair orientations*: percentages calculated from the counted puncta pairs in the previous columns Fig. 5 (tab 9): all data corresponding to Fig. 5 * *Embryo*: indicates biological replicate * *Cell Group Number*: analyzed cell identifier, corresponds to number of image in RawImages.zip, categorized based on orientation of the two WT:WT junctions (both vertical, both horizontal, or with one vertical and one horizontal junction) * *Number of cells in group:* the number of dual-reporter cells in the cluster of analyzed cells * *Junction orientation*: designates if the analyzed junction is vertical (between 45-90 degrees relative to the AP axis) or horizontal (between 0-45 degrees relative to the AP axis) * *Number of puncta pairs*: counted number of puncta pairs in either F:V, V:F or ambiguous orientations per WT:WT junction between dual-reporter cell groups in *Fz6-3xGFP; tdTomato-Vangl2 : Celsr1-/-* *chimeric skin * *Percentage of puncta pair orientations*: percentages calculated from the counted puncta pairs in the previous columns #### File: ads5704\_RawImages.zip **Description:** All raw .tif images of analyzed cell, cell pairs or groups of cells are organized into folders corresponding with the figure where analyses are presented. STED super-resolution images uploaded are deconvolved using NIS elements software. All cells, cell pairs, or cell groups are labeled with numbers that correspond with the numbers in the uploaded excel sheet, as described above. Files can be opened and analyzed using ImageJ, Tissue Analyzer or MATLAB. ## Code/software #### File: ads5704\_MATLAB scripts.zip **Description:** MATLAB scripts required to analyze PCP protein puncta pair orientations along junctions acquired with STED super-resolution imaging. Data processing and analysis using scripts detailed in materials and methods of this study.