Ajuba-GFP localization is spatiotemporally regulated and correlates with myosin. (A) Images from a time-lapse video of Ajuba-GFP during convergent extension. Stage 6 (−10 to 0 min), stage 7 (0–10 min), and stage 8 (10–30 min). (B) Ajuba-GFP intensity at cell interfaces of different orientations at t = 5 min). Angles are indicated relative to the AP axis, which is horizontal in all images (n = 6 embryos, 156–372 interfaces analyzed/embryo). (C) Image from a time-lapse video of Ajuba-GFP (green in merge) and Myo-mCherry (magenta in merge). (D) Myo-mCherry and Ajuba-GFP intensity are positively correlated during convergent extension. Each dot shows the mean Myo-mCherry and Ajuba-GFP intensity for a single interface, n = 190 interfaces in one embryo (R values from other embryos: 0.69, 0.72, 0.73, 0.77). (E) Kymograph of Ajuba-GFP (green) and Myo-mCherry (magenta) localization at a single cell interface. Yellow boxes highlight pulses of increased myosin and Ajuba localization. Arrowheads in E correspond to arrowheads in F. Images were acquired at 10-s intervals. (F) Mean Myo-mCherry and Ajuba-GFP intensity at the interface shown in E. (G) Correlation coefficient values between the change in Myo-mCherry interface intensity and the change in Ajuba-GFP (green line) or membrane-GFP (Resille-GFP, gray line). Ajuba-GFP and Resille-GFP data were shifted by the times shown on the x axis (n = 5 embryos/genotype, 10 edges analyzed/embryo). The mean ± SEM between embryos is shown in B and G. Images are anterior left, ventral down. Bars: (A and C) 10 µm; (E) 3 µm. See also Figs. S1 and S2.