Immobilisation and Wnt16 knockdown affects cell proliferation and migration at the medial region of the jaw joint between 3 and 5 dpf. (A-D′) Tg(Sox10:GAL4-VP16) and Tg(UAS:Kaede) transgenic line drives expression of Kaede protein (green) in the cartilage of control (A), anaesthetised (B), control-injected (C) and Wnt16 morpholino (MO)-injected (D) zebrafish. Maximum projections of the jaw joint from stacks of tiff images (A,B,C,D) and single slice/substacks through the same jaw joint to show cell morphology (A′,B′,C′,D′) are represented. At the jaw joint, medially located Kaede-expressing cells are photoconverted to red Kaede at 3 dpf (left panels). Right panels show jaw joints from the same larva re-imaged at 5 dpf. Photoconverted cells appear red/orange owing to presence of photoconverted red Kaede and expression of newly made green Kaede protein under control of the sox10 promoter. (E) Percentage increase in total area of cells expressing photoconverted red Kaede between 3 and 5 dpf in control, anaesthetised, control-injected and Wnt16 MO-injected zebrafish jaw joints (n=17, 18, 10 and 8 joints, respectively). (F) Percentage increase in number of cells expressing photoconverted red Kaede between 3 and 5 dpf in control, anaesthetised, control-injected and Wnt16 MO-injected zebrafish jaw joints (n=17, 16, 10 and 10 joints, respectively). Kruskal–Wallis tests were performed for E,F. ns, not significant, *P≤0.05, ***P≤0.001. Data are mean and 95%CI.
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