View largeDownload slide Laminins perform multiple roles during zebrafish heart morphogenesis. (A-F) mRNA in situ hybridisation analysis of myl7 expression in control embryos injected with lamc1-targeting gRNAs only (A,C,E) or with lamc1-targeting gRNAs together with Cas9 protein (lamc1 F0; B,D,F) at 30 hpf (A,B), 55 hpf (C,D) and 72 hpf (E,F). (G-L) Quantitative analysis of looping ratio (G,I,K) and myl7 area (H,J,L) in gRNA-injected controls (30 hpf: n=34; 55 hpf: n=44; 72 hpf: n=44) and lamc1 F0 crispants (30 hpf: n=38; 55 hpf: n=47; 72 hpf: n=44). lamc1 crispants exhibit reduced heart looping at 55 hpf and 72 hpf, a reduced area of myl7 expression at 30 hpf and an increased area of myl7 expression at 72 hpf. Data are median±interquartile range, analysed with the Kruskal–Wallis test. (M-R) mRNA in situ hybridisation analysis of myl7 expression in siblings (M,O,Q) and lamb1aΔ25 mutants (N,P,R) at 30 hpf, 55 hpf and 72 hpf. (S-X) Quantitative analysis of looping ratio (S,U,W) and myl7 area (T,V,X) in siblings (30 hpf: n=65; 55 hpf: n=70; 72 hpf: n=56) and lamb1aΔ25 mutants (30 hpf: n=20; 55 hpf: n=25; 72 hpf: n=34). lamb1aΔ25 mutants exhibit a mild reduction in heart looping from 55 hpf, and an increased area of myl7 expression at 55 hpf and 72 hpf. Data are median±interquartile range, S-W were analysed with the Mann–Whitney U test, X was analysed with the Kruskal-Wallis test. ****P<0.0001, ***P<0.001, **P<0.01, ns=not significant in all graphs. Scale bars: 50 μm.
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