jnk3 is required to restrict pitx2c. (A) Representative bright-field image of MZjnk3 mutant at 3 dpf. (B) Representative MZjnk3 KV at 10 ss labelling acetylated tubulin (green) and aPKC (magenta). (C) Quantification of number of cilia in wild-type and MZjnk3 embryos at 10 ss. Loss of jnk3 results in a significant increase in the number of nodal cilia. (D) Quantification of length of nodal cilial in wild-type and MZjnk3 embryos at 10 ss. Loss of jnk3 does not affect cilial length. (E,E′) Quantification of nodal flow speed in MZjnk3 by quadrant (µm/s; data are mean±s.d.) (E) and average speed (E′) between 10 and 14 ss. Loss of jnk3 results in a significant reduction in average speed (E′). (F) Loss of jnk3 does not impact normal spaw expression at 12-14 ss. (G) Characterisation of abnormal pitx2c expression in wild type and MZjnk3 mutants at 18-19 ss. Loss of jnk3 leads to a significant increase in the proportion of embryos that display bilateral pitx2c expression. (H) Heart jogging is unaffected in MZjnk3 mutants. (I) MZjnk3 mutants have a significant proportion of bilaterally positioned abdominal organs. (C) Data are mean±s.d., Welch's t-test. MZjnk3, n=14. Wild-type data are from Fig. 1F. (D) Data are mean±s.d., Welch's t-test. MZjnk3, n=720. Wild-type data are from Fig. 1H. (E′) Data are median±interquartile range, Mann–Whitney test. MZjnk3, n=61 beads across 11 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: MZjnk3, n=23. 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: MZjnk3, 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=8 clutches for MZjnk3. Minimum clutch size: MZjnk3, n=46. Wild-type data are from Fig. 4H. (I) Data are mean±s.e.m., two-way ANOVA comparison of right and bilateral. n=3 clutches. Minimum clutch size: MZjnk3, n=92. Wild-type data are from Fig. 4I. (A) Lateral view, anterior leftwards. Scale bar: 1 mm. (B) Anterior upwards. Scale bar: 20 µm. ns, not significant. *P<0.05, **P<0.01, ****P<0.0001. AL, anterior left; AR, anterior right; PL, posterior left; PR, posterior right.
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