jnk1a and jnk1b function redundantly to regulate cilia length in the zebrafish left-right organiser. (A) Schematic of zebrafish embryo at the 10-somite stage (ss), lateral view. Kupffer's vesicle (KV, magenta) is located at the caudal tip of the notochord. The caudal region of the embryo imaged in B-E is outlined. (B,B′) Representative image of wild-type KV at 10 ss used for characterisation of cilial parameters by immunohistochemistry for acetylated tubulin with depiction of anterior (blue) and posterior (yellow) in KV (B) and aPKC (B′). (B″) Merged images. (C-E) Representative MZjnk1a (C), MZjnk1b (D) and MZjnk1a;MZjnk1b (E) KVs at 10 ss labelling acetylated tubulin (green) and aPKC (magenta). (F) Quantification of the number of cilia in KV in wild-type, MZjnk1a, MZjnk1b and MZjnk1a;MZjnk1b embryos at 10 ss. jnk1 mutants display no differences in the number of nodal cilia in KV. (G) Quantification of nodal cilia distribution in wild-type, MZjnk1a, MZjnk1b and MZjnk1a;MZjnk1b embryos at 10 ss. Antero-posterior distribution nodal cilia is unaffected in jnk1 mutants. (H) Quantification of length of nodal cilial in wild-type, MZjnk1a, MZjnk1b and MZjnk1a;MZjnk1b embryos at 10 ss. MZjnk1a and MZjnk1b mutant embryos display a similar significant reduction in cilia length. Loss of both jnk1a and jnk1b results in a more dramatic reduction in cilia length. (F) Data are mean±s.d., one-way ANOVA, multiple comparisons. Wild type, n=17; MZjnk1a, MZjnk1b and MZjnk1a;MZjnk1b, n=16. (G) Data are mean±s.e.m., two-way ANOVA, multiple comparisons, n=16. (H) Data are mean±s.d., one-way ANOVA, multiple comparisons. Wild type, n=809; MZjnk1a, n=812; MZjnk1b, n=743; MZjnk1a;MZjnk1b, n=754. ns, not significant; *P<0.05, ****P<0.0001. (B-E) Anterior is upwards, left is leftwards. Scale bars: 20 µm.
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