The M1–Vinc interaction supports Ecad stability and reinforces adhesion. (A) Ecad planar polarity (enrichment at vertical edges) is shown in α-Cat-RNAi α-CatX embryos at stage 8 expressing either GAP43::mCherry or Vinc::mCherry. n = number of embryos (e). Significance determined by ordinary two-way ANOVA with uncorrected Fisher’s LSD. (B) An extra copy of Vinc (Vinc::mCherry) improves adhesion in the ectoderm of α-Cat-RNAi α-Cat-ΔM23 but not α-Cat-RNAi α-Cat-ΔM embryos at stage 8 (live imaged stills shown). Quantification of area of gaps (orange shading) at right. Significance given by Mann-Whitney two-tailed test, n = number of embryos (e). (C and D) Comparison of control and Vinc-CO embryos expressing Ecad::GFP. Stills from live embryos at stage 8. FI of Ecad and Ecad planar polarity (D) shown, n = number of embryos (e). For A, B, and D, bar height represents mean and error bars show SD. (E) Denticle belt count of α-Cat-RNAi α-Cat-ΔM23 embryos with and without a Vinc maternal-zygotic (MZ) null mutant background. Number of embryos analyzed = n, significance determined by Chi-square test. (F) Live imaged stills comparing Jub::GFP signal in wild-type and a Vinc MZ null mutant embryo at stage 8. FI of Jub::GFP in these conditions shown. (G)α-Cat-RNAi α-CatR-ΔM1 embryos at stage 8 show less uniform junctional distribution of Ecad::GFP compared to α-Cat-RNAi α-CatR controls as indicated by the increased normalized SD of Ecad::GFP signal at AJs. (H) Junctional α-Cat::YFP signal (same color map in F) in stage 8 embryos is reduced and less uniformly distributed in the absence of Vinc, as indicated by an increase of the normalized SD of α-Cat::YFP signal. Fluorescent intensity in F and H displayed as in color map shown, maximum intensity (4095), white. For B, C, F, G, and H, scale bar, 10 μm. For C, F, G, and H, bold line shows the median, dotted lines represent interquartile ranges, and n = a pooled number of cells (c) or junctions (j) for a number of embryos (e). Mann-Whitney two-tailed test was used to determine significance for C, D, F, G, and H, **** = P < 0.0001, *** = P < 0.001, ** = P < 0.01, * = P < 0.05. (I) Schematic summary: Vinc does not play an inhibitory role at AJs. Recruitment of Vinc to the junction depends on M1, and this supports Ecad at higher-tension vertical edges and adhesion. This is in parallel to M2/3, which also supports Ecad recruitment to vertical edges.
Figure 6.

The M1–Vinc interaction supports Ecad stability and reinforces adhesion. (A) Ecad planar polarity (enrichment at vertical edges) is shown in α-Cat-RNAi α-CatX embryos at stage 8 expressing either GAP43::mCherry or Vinc::mCherry. n = number of embryos (e). Significance determined by ordinary two-way ANOVA with uncorrected Fisher’s LSD. (B) An extra copy of Vinc (Vinc::mCherry) improves adhesion in the ectoderm of α-Cat-RNAi α-Cat-ΔM23 but not α-Cat-RNAi α-Cat-ΔM embryos at stage 8 (live imaged stills shown). Quantification of area of gaps (orange shading) at right. Significance given by Mann-Whitney two-tailed test, n = number of embryos (e). (C and D) Comparison of control and Vinc-CO embryos expressing Ecad::GFP. Stills from live embryos at stage 8. FI of Ecad and Ecad planar polarity (D) shown, n = number of embryos (e). For A, B, and D, bar height represents mean and error bars show SD. (E) Denticle belt count of α-Cat-RNAi α-Cat-ΔM23 embryos with and without a Vinc maternal-zygotic (MZ) null mutant background. Number of embryos analyzed = n, significance determined by Chi-square test. (F) Live imaged stills comparing Jub::GFP signal in wild-type and a Vinc MZ null mutant embryo at stage 8. FI of Jub::GFP in these conditions shown. (G)α-Cat-RNAi α-CatR-ΔM1 embryos at stage 8 show less uniform junctional distribution of Ecad::GFP compared to α-Cat-RNAi α-CatR controls as indicated by the increased normalized SD of Ecad::GFP signal at AJs. (H) Junctional α-Cat::YFP signal (same color map in F) in stage 8 embryos is reduced and less uniformly distributed in the absence of Vinc, as indicated by an increase of the normalized SD of α-Cat::YFP signal. Fluorescent intensity in F and H displayed as in color map shown, maximum intensity (4095), white. For B, C, F, G, and H, scale bar, 10 μm. For C, F, G, and H, bold line shows the median, dotted lines represent interquartile ranges, and n = a pooled number of cells (c) or junctions (j) for a number of embryos (e). Mann-Whitney two-tailed test was used to determine significance for C, D, F, G, and H, **** = P < 0.0001, *** = P < 0.001, ** = P < 0.01, * = P < 0.05. (I) Schematic summary: Vinc does not play an inhibitory role at AJs. Recruitment of Vinc to the junction depends on M1, and this supports Ecad at higher-tension vertical edges and adhesion. This is in parallel to M2/3, which also supports Ecad recruitment to vertical edges.

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