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. 2022 Nov 18;33(14):ar136. doi: 10.1091/mbc.E22-06-0205

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FIGURE 4: — p115RhoGEF prevents repeated local leaks to maintain TJ barrier function. (A) Time-lapse images (Fire LUT) of whole-field epithelial barrier permeability determined using ZnUMBA (FluoZin-3 intensity) in control and p115.S KD embryos. Time 0 represents mounting of embryos in 1 mM Zinc-chloride in 0.1 × MMR and the start of the time-lapse movie. (B) Graph of mean normalized intensity of whole-field FluoZin-3 signal intensity in control (black line) and p115.S KD (green line) embryos. Whole-field FluoZin-3 signal increases rapidly over time upon p115.S KD in comparison to control. Shading represents SEM. Control: n = 5 embryos, 3 experiments; p115.S KD: n = 5 embryos, 3 experiments. (C, D) Cell view of FluoZin-3 signal in control (C) and p115.S KD (D) embryos. White dashed rectangles represent the junctions used to make the kymographs. Kymographs of FluoZin-3 (green), occludin (mCherry-occludin, magenta), and merged image projected from vertex-to-vertex over time in control and p115.S KD embryos. Control embryos display one prominent FluoZin-3 leak (asterisk) and occludin reinforcement (white bracket). Repeated local barrier leaks (white asterisks) and lack of occludin reinforcement occur in p115.S KD embryos.