Xenbase Image ID: 158559

Supplementary Figure 2 | Experimental manipulation adhesion and actomyosin contractility. (a) Cadherin overexpression and depletion. Comparison by immunoblot of C-cadherin protein levels in control tissues and tissues from embryos injected with C-cadherin-GFP mRNA or C-Cadherin MO. Arrow, endogenous C-cadherin; arrowhead: Ccadherin-GFP. Quantification was performed from three independent experiments. Individual comparisons were done using Games-Howell test (* P<0.05) after a significant One-way Welch ANOVA (P<0.0001; unequal variances). Error bars are s.d. (b) Validation of cadherin depletion and replacement. C-cadherin was depleted by MO injection and replaced with E- or N-cadherin by co-injection of the corresponding mRNA. GFP mRNA was co-injected as a tracer. Manipulated ectoderm explants were juxtaposed to wild type non-injected ectoderm explants for direct comparison of β-catenin membrane immunostaining, used as a general marker for cadherin-based adhesive structures. Pseudocolors were added to help visualizing the decrease of β-catenin signal caused by CadMO and the rescue by expression of E- or N-cadherin. Numbers of samples indicated at the bottom. Scale bar, 30μm. (c) Effect of C-cadherin overexpression, depletion, and replacement on ectoderm and mesoderm cohesion. The same conditions as in (b) were tested for resistance to dissociation. Graph shows mean values and error bars are s.d. Number of experiments is given above the graph. Individual comparisons were done using Games-Howell test (* P<0.05) after a significant one-way Welch ANOVA P<0.0001; for unequal variances). (d) Manipulation of cortical contractility. Top left panel: Effect of MHC2A+B depletion (MHCMOs) and Rho activation (caRhoA mRNA) on the elastic modulus of ectoderm and induced mesoderm cells. Data are from 5 independent experiments. Right: Effect of Eph stimulation by soluble ephrinB3-Fc ligand. Lower left: Single myosin isoform depletion (MHC2A or MHC2B MO) did not cause a decrease in elastic modulus, suggesting that each isoform is able to compensate for the loss of the other. Box plots as in main Fig.2e. The number of cells tested/independent experiment is above the graph. Individual comparisons were done using Tukey’s Hs.d test after a significant One-way ANOVA (P=5.7e-08). Image published in: Canty L et al. (2017) © The Author(s) 2017. 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

Return to previous page