jnk2 regulates nodal cilia development and KV flow. (A) Representative bright-field image of MZjnk2 mutant at 3 dpf. (B) Representative MZjnk2 KV at 10 ss labelling acetylated tubulin (green) and aPKC (magenta). (C) Quantification of number of cilia in wild-type and MZjnk2 embryos at 10 ss. Loss of jnk2 results in a significant increase in the number of nodal cilia. (D) Quantification of length of nodal cilial in wild-type and MZjnk2 embryos at 10 ss. MZjnk2 mutant embryos have a significant reduction in the length of nodal cilia. (E,E′) Quantification of nodal flow speed in MZjnk2 by quadrant (μm/s; data are mean±s.d.) (E) and average speed (E′) between 10 and 14 ss. Loss of jnk2 results in a significant reduction in average speed (E′). (F) Loss of jnk2 does not impact normal spaw expression at 12-14 ss. (G) Characterisation of abnormal pitx2c expression in wild type and MZjnk2 mutants at 18-19 ss. Loss of jnk2 results in a significant proportion of embryos that do not have pitx2c expression in the lateral plate mesoderm. (H) Heart jogging is unaffected in MZjnk2 mutants. (I) Gut looping occurs normally in MZjnk2 mutants. (C) Data are mean±s.d., Welch's t-test. MZjnk2, n=14. Wild-type data are from Fig. 1F. (D) Data are mean±s.d., Welch's t-test. MZjnk2, n=737. Wild-type data are from Fig. 1H. (E′) Data are median±interquartile range, Mann–Whitney test. MZjnk2, n=28 beads from six embryos. Wild-type data are from Fig. 2F. (F) Data are mean±s.e.m., two-way ANOVA comparison of right and bilateral. n=3 clutches. Minimum clutch size: MZjnk2, n=24. Wild-type data are from Fig. 3B. (G) Data are mean±s.e.m., two-way ANOVA, multiple comparisons analysing right, bilateral and absent. n=5 clutches. Minimum clutch size: MZjnk2, n=81. Wild-type data are from Fig. 3H. (H) Data are mean±s.e.m., two-way ANOVA comparison of right and no jog. n=6 clutches for wild type and MZjnk2. Minimum clutch sizes: wild type, n=101; MZjnk2, n=46. (I) Data are mean±s.e.m., two-way ANOVA comparison of right and bilateral. n=3 clutches. Minimum clutch sizes: wild type, n=98; MZjnk2, n=89. (A) Lateral view, anterior leftwards. Scale bar: 1 mm. (B) Anterior upwards. Scale bar: 20 µm. ns, not significant. *P<0.05, ****P<0.0001. AL, anterior left, AR, anterior right; PL, posterior left; PR, posterior right.
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