|
Fig. S1
A second non-overlapping morpholino, against the exon3-intron3 splice site (fto spl.MO), confirms specificity of the fto phenotype. (A) fto spl. morphants show a similar general morphology to fto ATG morphants, displaying small eyes, reduced pharyngeal length, and curved truncated body axis at 48 hpf and 5 dpf. Scale bar: 500 μm. (B) Craniofacial defects were also observed in the fto spl. morphants, as in fto ATG morphants, assayed using alcian blue to detect cartilage. Scale bar: 200 µm. (C) RT-PCR of a product surrounding the E3-I3 splice site confirmed fto knockdown and specificity of the fto Spl.MO at 48 hpf, presumably due to the two in-frame intronic stop codons, 72 nt and 84 nt into intron 3, causing RNA mediated decay. GAPDH was used as a loading control.