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Figure 7. C-cadherin is required for endoderm cell migration.(A) Protrusions engage in cell-cell adhesion. Labelled (mRFP) endoderm cells are elongated (yellow outline) in explants (left). Cells project protrusions (yellow arrows) onto neighbouring cells (center). Protrusions are enriched with α-catenin (blue arrow) at sites of cell-cell contact (right). (B) C-cadherin knockdown altered cell morphology and reduced protrusion formation. C-cadherin morpholino (CcadMO) injected cells appear rotund (pink outline) (left) and extend few protrusions (center). However, α-catenin (blue arrow) was still present within protrusions (right). Region of interest (red box) is indicated in the top right corners. (C) Cell length-width ratio in uninjected and CcadMO-injected explants. Cells were sampled from the middle of the central column of the endoderm. (D) Cell elongation congruity in uninjected and CcadMO-injected explants. For C and D, plots show data sampled from three embryos from different egg batches. (E) Average number of protrusions per cell in uninjected and CcadMO-injected explants. For all histograms, error bars indicate S.E. (F, G). Protrusion dynamics of uninjected and CcadMO-injected cells. Zero indicates no change (dash grey line), net gain (positive axis) and net loss (negative axis) of protrusions between consecutive time points are shown. Colours represent individual cells. For E–G, plots show data from three embryos from different egg batches. (H) Quantification of tissue cohesion. Tissue surface tension measured from uninjected and CcadMO-injected endoderm. Plot shows data from 22 embryos from different egg batches. (I) Cell migration velocity in explants comparing uninjected and CcadMO-injected cells. (J, K) Movement trajectories of cells in explants. (J) Uninjected, (K) CcadMO-injected explant after 60 min. Colours represent individual cells. For I–J, plots show data from four embryos collected from different egg batches.10.7554/eLife.27190.024Figure 7—source data 1. Quantification of the effects of C-cadherin knockdown on cell morphology and behaviour.Figure 7—figure supplement 1. Localization of α-catenin.(A) Localization of α-catenin-GFP at sites of cell–cell contact (white arrows). (B) Protrusion extension is correlated with α-catenin accumulation (top row, white arrows). α-catenin enrichment fills protrusions from the tip to the base (bottom row). (C) Protrusion retraction is correlated with α-catenin reduction (white arrows). After complete retraction, α-catenin is diminished in the cortex (bottom row). Image published in: Wen JW and Winklbauer R (2017) © 2017, Wen et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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