Fig. 2
Loss of ik results in abnormal ciliary morphology (A) Whole-mount immunostaining of cilia in the inner ear (white boxes) using anti-acetylated-α-tubulin (green) of WT embryos (n = 3) and ik mutants (n = 5) at 2 dpf. Scale bar, 20 μm. Quantified graphs of cilia lengths and numbers in the inner ear between WT embryos and ik mutants at 2 dpf. *p < 0.05, ***p < 0.001 (B) Confocal microscopic images of control MO, ik MO, and ik MO/ik mRNA co-injected Tg(brn3c:GFP) embryos at 3 dpf. Scale bar, 20 µm. Quantification of ciliary length and number in the inner ear of control MO (n = 12), ik MO (n = 17), and ik MO/ik mRNA co-injected embryos (n = 10). **p < 0.01, ***p < 0.001 (C) Whole-mount immunofluorescence (left) of overall cilia in the pronephros in WT and in ik mutants at 2 dpf using anti-acetylated-α-tubulin (green). Scale bar, 100 µm. Enlarged view (right) of pronephric cilia in proximal and distal tubules stained with anti-acetylated-α-tubulin (green) and nuclei stained with DAPI (blue) at 2 dpf WT embryos and ik mutants. Scale bar, 20 µm. Stacked bar graph displays the percentage of embryos of cilia phenotype in the anterior pronephric duct of WT embryos and ik mutants at 2 dpf. (D) TEM results showing the ultrastructure of cilia in the pronephric duct of ik mutants. Cross-section showing the “9 + 2” configuration (black circles). Scale bar, 100 nm (E) Whole-mount immunofluorescence (left) of overall cilia in the pronephros of control MO, ik MO, and ik MO/ik mRNA co-injected embryos at 3 dpf with anti-acetylated-α-tubulin (green). Scale bar, 100 µm. Enlarged view (right) of pronephric cilia in the anterior and posterior pronephric ducts stained using anti-acetylated-α-tubulin (green) and nuclei stained with DAPI (blue) at 2 dpf control MO, ik MO, and ik MO/ ik mRNA co-injected embryos. Scale bar, 20 µm. Stacked bar graph displays the percentage of embryos based on ciliary morphology in the anterior pronephric duct (percentage of abnormal ciliary morphology of embryos: control MO, 0%; ik MO, 58%; ik MO + ik mRNA, 0%)