M region supports mechanosensitive recruitment of Jub to AJs. (A) Stills from live α-Cat-RNAi α-CatX embryos expressing Jub::GFP. FI is displayed as shown in color map at top. Maximum FI (255), white. Scale bar, 10 μm. (B and C) The mean FI of Jub (with background cytoplasmic signal subtracted) for α-Cat-RNAi α-CatX embryos at stage 8 is plotted for (B) horizontal edges (0–15°) and (C) vertical edges (75–90°; 90° = DV axis). n = number of embryos (e); bold line, median; dotted lines, interquartile ranges. Significance determined by ordinary one-way ANOVA with Tukey’s multiple comparisons test. (D) Mean FI of Vinc, Jub, Cno at horizontal BCJs, vertical BCJs, and TCJs in stage 8 embryos. Points represent mean FI of regions of interest per embryo (for Vinc, n = 9; Jub, n = 9; Cno n = 4). (E) Quantification of planar polarity of Jub is plotted per embryo as mean FI of vertical edges divided by that of horizontal edges, derived from data in B and C. Significance determined by ordinary two-way ANOVA with uncorrected Fisher’s LSD for all bins. See Fig. S3 for more detailed breakdown of enrichment of FI by angles of edges. (F) Quantification of enrichment of Jub signal at TCJs vs. BCJs within the same embryo. Significance determined by ordinary one-way ANOVA with Tukey’s multiple comparison test. For D, E, and F, height of bars represents mean, and error bars represent SD. (G) Live-imaged stills of α-Cat-RNAi α-CatX embryos expressing Jub::GFP, with close-ups highlighting TCJs (orange arrowheads). Scale bar, 10 μm; color map same as in A. (H) Analysis of cuticle defects of α-Cat-RNAi α-CatX embryos and either GFP-RNAi as a control or Jub-RNAi. Number of embryos analyzed = n. Significance determined by Chi-square test. For B, C, E, F, and H, **** = P < 0.0001, *** = P < 0.001, ** = P < 0.01, * = P < 0.05. (I) Schematic summary: Junctional recruitment of Jub depends on the interaction between Jub and the N2 domain. This interaction is inhibited at horizontal edges by M1 and is supported at higher tension vertical edges by M2, and at TCJs by M2 and M3.
Figure 5.

M region supports mechanosensitive recruitment of Jub to AJs. (A) Stills from live α-Cat-RNAi α-CatX embryos expressing Jub::GFP. FI is displayed as shown in color map at top. Maximum FI (255), white. Scale bar, 10 μm. (B and C) The mean FI of Jub (with background cytoplasmic signal subtracted) for α-Cat-RNAi α-CatX embryos at stage 8 is plotted for (B) horizontal edges (0–15°) and (C) vertical edges (75–90°; 90° = DV axis). n = number of embryos (e); bold line, median; dotted lines, interquartile ranges. Significance determined by ordinary one-way ANOVA with Tukey’s multiple comparisons test. (D) Mean FI of Vinc, Jub, Cno at horizontal BCJs, vertical BCJs, and TCJs in stage 8 embryos. Points represent mean FI of regions of interest per embryo (for Vinc, n = 9; Jub, n = 9; Cno n = 4). (E) Quantification of planar polarity of Jub is plotted per embryo as mean FI of vertical edges divided by that of horizontal edges, derived from data in B and C. Significance determined by ordinary two-way ANOVA with uncorrected Fisher’s LSD for all bins. See Fig. S3 for more detailed breakdown of enrichment of FI by angles of edges. (F) Quantification of enrichment of Jub signal at TCJs vs. BCJs within the same embryo. Significance determined by ordinary one-way ANOVA with Tukey’s multiple comparison test. For D, E, and F, height of bars represents mean, and error bars represent SD. (G) Live-imaged stills of α-Cat-RNAi α-CatX embryos expressing Jub::GFP, with close-ups highlighting TCJs (orange arrowheads). Scale bar, 10 μm; color map same as in A. (H) Analysis of cuticle defects of α-Cat-RNAi α-CatX embryos and either GFP-RNAi as a control or Jub-RNAi. Number of embryos analyzed = n. Significance determined by Chi-square test. For B, C, E, F, and H, **** = P < 0.0001, *** = P < 0.001, ** = P < 0.01, * = P < 0.05. (I) Schematic summary: Junctional recruitment of Jub depends on the interaction between Jub and the N2 domain. This interaction is inhibited at horizontal edges by M1 and is supported at higher tension vertical edges by M2, and at TCJs by M2 and M3.

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