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Knockdown phenotype of fibronectin in S1PRs mutant. (A) Membrane topology of S1P receptors and their mutants. The region of frameshift-mediated amino acids compared with the S1PR2 wild type (WT) is shown by the dashed line. (B,C,D) Fluorescence microscopy of intersegmental vessels of wt (B), s1pr1 mutant (C) and s1pr2 mutnat (D) at 2dpf. Endothelial cells are visualized by EGFP expression derived from Tg(fli1a:EGFP). (E,F) Cardiac morphology visualized by mRFP expression derived from Tg(cmlc2:mRFP). All images show ventral views at 28hpf. (G) Average distances between two hearts from multiple experiments; error bars represent standard deviations. (H,I) Lower jaw morphology at 4dpf was visualized by Alcian Blue staining (ventral view). (J) Average anterior–posterior distances of the ventral pharyngeal arch from multiple experiments; error bars represent standard deviations. Genotyping was performed by genomic sequencing or heteroduplex mobility assays after taking pictures. Scale bars: 200μm.
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