Ajuba localization is sensitive to mechanical tension. (A) Ajuba-GFP and Myo-GFP localization in stage 7/8 embryos injected with water (left) or the Rho-kinase inhibitor Y-27632 (right). (B) Ajuba-GFP and Myo-mCherry in stage 8 WT (left) and Shroom mutant embryos (the progeny of Shroom^Δ11/Df males and females; right). The accumulation of Ajuba and myosin at vertical interfaces is lost in Y-27632–injected and Shroom mutant embryos. (C) Ajuba-GFP and Myo-mCherry in stage 6 WT and ShroomA-overexpressing (Shroom OE) embryos. Overexpression of ShroomA recruits Ajuba and myosin to cell vertices. (D) Ajuba-GFP enrichment at vertical edges (oriented at 75–90° relative to the AP axis) and horizontal edges (oriented at 0–15° relative to the AP axis) in stage 7/8 water- or Y-27632–injected embryos (n = 5 embryos/condition, 70–99 edges analyzed/embryo). (E) Ajuba-GFP enrichment at vertical and horizontal edges in stage 8 WT and Shroom mutant embryos (n = 5 embryos/genotype, 106–283 edges analyzed/embryo). Edge intensities were divided by the cytoplasmic intensity to calculate the junctional enrichment. (F) Ajuba-GFP vertex enrichment in stage 6 WT and Shroom OE embryos (n = 5 embryos/genotype, 30 vertices analyzed/embryo). Vertex intensities were divided by the cytoplasmic intensity to calculate the vertex enrichment. (G) Ajuba-GFP enrichment at all edges in stage 7 WT (gray circles) and Shroom OE embryos (white circles; n = 5 embryos/genotype, 75–102 edges analyzed/embryo). The mean ± SEM between embryos is shown in D–F; each dot in G shows the mean ± SD for a single embryo; *, P < 0.01; **, P < 0.001, unpaired Student’s t test. Images are anterior left, ventral down. Bars, 10 µm. See also Fig. S1.