Figure 7

Figure 7

vangl2 mutants have misshapen OFT. (A–B′) Representative brightfield image of WT sibling (A, n = 7) and vangl2^m209 homozygous mutant (B, n = 15) at 70 hpf, dashed boxes in (A) and (B) highlights heart shown in (A′) and (B′). The overt morphology of the vangl2^m209 OFT appears normal (white). (C and D) Representative images of mRNA in situ hybridization for elnb in sibling (C) and vangl2^m209 homozygous mutant (D) at 70 hpf. (E) Quantification of area of elnb domain at 70 hpf. Loss of vangl2 results in a larger elnb domain. (F) Quantification of roundness of elnb domain at 70 hpf. Loss of vangl2 leads to a less round elnb domain (WT sibling, n = 19; vangl2^m209, n = 28). (G and H) Representative midline sections of immunohistochemistry on WT sibling (G) and vangl2^m209 homozygous mutants (H) carrying Tg(kdrl:HsHRAS-mCherry) to mark the endocardium (green) and Tg(myl7:GFP) to mark the myocardium (cyan) and MLCK (magenta). The vangl2 OFT appears squat, with the myocardial collar less pronounced. (I and I′) Quantification of number of cells in arterial valve primordia (yellow) at 70 hpf in WT sibling and vangl2^m209 homozygous mutants at 70 hpf. Loss of vangl2 does not impact number of VICs ((WT sibling, n = 17; vangl2^m209, n = 15), D, dextral primordia; S, sinistral primordia). (J–J″) Quantification of arterial valve primordia volume by 3D reconstruction of WT sibling (J) and vangl2^m209 (J′) homozygous mutant OFT from immunohistochemistry. Loss of vangl2 does not impact primordia volume (J″). (E), (F), (I), and (I′): Mean ± SEM, Welch’s unpaired t-tests. (J″): Mean ± SEM, Brown–Forsyth and Welch’s ANOVA. ***, P < 0.001; ns, not significant. V, ventricle; BA, bulbus arteriosus; D, dextral; S, sinistral. Scale bars: (A) and (B): 500 μm; (A′) and (B′): 50 μm; (C), (D), (J), and (J′): 20 μm; (G) and H): 10 μm.