Fig. 5

Zfcobl morphants exhibit reduced motile cilia length.

Anti-acetylated tubulin staining reveals shortened cilia in Kupffer′s vesicle at eight somites in control (A) and cobl morphant (B) embryos. (C) Bar graph showing mean cilia length in microns (μm). Control embryos have a mean cilia length of 6.75 μm in Kupffer′s vesicle. KV cilia in cobl morphants show an average reduction of 20–50% in length. n values: controls = 694 cilia, MOa(6 ng) = 2011, MOa(3 ng) = 738, MOb(16 ng) = 579, MOa + MOb = 574. p value: C = < 0.0001* (each pair). (D–E) Anti-acetylated tubulin staining reveals reduced cilia length in the pronephros at 24 hpf in cobl morphants. (F) Bar graph showing mean cilia length in microns. n values: controls = 167cilia, MOa = 153. p value: F = < 0.0001*. (G–H) Anti-acetylated tubulin staining reveals reduced cilia length on multi-ciliated cells in the pronephros at 48 hpf. (I) Bar graph showing mean multi-cilia length in microns. n values: controls = 19, MOa = 20. p value: I = < 0.0001*. Cilia number is unaffected in cobl morphants relative to controls. (J) Bar graph depicting average cilia number in Kupffer′s vesicle in 8-somite control embryos vs. cobl morphant embryos. No significant difference is observed between controls and the various morphant groups. (K) Bar graph depicting average cilia number in the pronephric ducts in 24 hpf control embryos vs. cobl-MOa embryos. Z-stacks were taken through the middle region of the pronephric duct (∼ 141 μm length frame). Cilia were stained with anti-aTub and anti-gTub and counted. Error bars represent one standard error from the mean. p values for controls versus morphants all > 0.05. Bar with asterisk highlights specific significance. Arrows in A–B, D–E, G–H point to cilia. Scale bars represent 10 μm.