Asymmetric spaw expression is dispensable for organ asymmetry. (A) Characterisation of abnormal spaw expression in wild type and MZjnk1a;MZjnk1b;Zjnk2 mutants at 12-14 ss. Loss of jnk1 and jnk2 results in ∼40% of embryos displaying either right or bilateral spaw expression. (B) Characterisation of abnormal pitx2c expression in wild type and MZjnk1a;MZjnk1b;Zjnk2 mutants at 18-19 ss; loss of jnk1 and jnk2 results in a significant increase in the proportion of embryos without pitx2c expression in the LPM. (C-C″) Representative images of mRNA in situ hybridisation for the pan-cardiac marker myl7 at 72 hpf showing normal dextral looping of the heart (C), abnormal sinistral (reverse) looping (C′) or non-looped hearts (C″). (D,E) Quantification of (D) heart looping and (E) gut looping in wild type and MZjnk1a;MZjnk1b;Zjnk2 mutants at 72 hpf. Loss of jnk1 and jnk2 activity does not impact organ laterality. (A) Data are mean±s.e.m., two-way ANOVA comparison of right and bilateral. n=3 clutches. Minimum clutch size: MZjnk1a;MZjnk1b;Zjnk2, n=10. Wild-type data are from Fig. 3B. (B) Data are mean±s.e.m., two-way ANOVA, multiple comparisons analysing right, bilateral and absent. n=5 clutches. Minimum clutch size: MZjnk1a;MZjnk1b;Zjnk2, n=40. Wild-type data are from Fig. 3H. (D) Data are mean±s.e.m., two-way ANOVA comparison of sinistral and no loop. n=3 clutches. Minimum clutch sizes: wild type, n=88; MZjnk1a;MZjnk1b;Zjnk2, n=34. (E) Data are mean±s.e.m., two-way ANOVA comparison of right and bilateral. n=3 clutches, same clutches as in D. (C-C″) Ventral view. ns, not significant. *P<0.05, **P<0.01. V, ventricle; A, atrium.
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