Fig. S1

Fig. S1 Aoh is a furina mutant. Laterality phenotypes of Zaoh and MZaoh. (A) Anterior and posterior expression of laterality markers in wt, Zaoh and MZaoh embryos. Reduced but maintained expression of pitx2c in Zaoh is indicated by arrowhead. (B) Genomic position of the aoh mutation. (C) A single nucleotide substitution in furinA (G to A) resulting in a tryptophan (Trp) to stop (TAG) mutation at amino acid 411 is responsible for the aoh mutation. (D) Complementation test. Outcross of aoh^+/- to stu^+/- (Walker et al., 2006) fails to complement the aoh cardiac jogging phenotype. Stacked histograms display averages +/- SEM collected over 3 independent biological replicates. (E) FurinA is maternally contributed to the embryo and is broadly expressed during somatogenesis in wt and sfw/spaw^-/- embryos. (F) Addition of FurinA mRNA to Spaw-eGFP mRNA injected in 1-cell embryos results in significantly reduced Spaw-eGFP precursor (*) in 12-somite stage embryos. Whole embryo extracts were subjected to immunoblotting with the indicated antibodies. Actin is shown as loading control. Mature Spaw-eGFP (**) appears as a faint band of approximately 41 kDa. Quantification of the SpaweGFP precursor abundance is depicted in the histogram. *** P<0.001 (one-way ANOVA). Data collected over three biologically and technically independent replicates.