DEV011015 Supplementary Material

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• Supplemental Figure S1 -

  Fig. S1. Co-injection of zebrafish embryos with morpholino antisense oligos (MOs) and the hybrid RNA encoding GFP fused to the 5′ untranslated and initial 20-bp coding region of their target genes. (A) Schematic of experimental procedures. The hybrid RNA or a mixture of the hybrid RNA and MOs was injected into one-cell stage embryos. The hybrid RNA activates GFP expression a couple of hours after injection (above), but simultaneous introduction of MOs represses this RNA-mediated GFP expression if MOs specifically inhibit the translation of their target genes (beneath). We examined GFP fluorescence in embryos injected with different combinations of RNAs and Mos: (B) no RNA and no MO; (C) the hybrid RNA encoding GFP fused to the 5′ untranslated and initial 20-bp coding region of the chk1 gene RNA(Chk1:GFP) and no MO; (D) RNA(Chk2:GFP) and no MO; (E) RNA(Chk1:GFP) and MO-Chk1; (F) RNA(Chk2:GFP) and MO-Chk2; (G) RNA(Chk1:GFP) and 5mis-MO-Chk1; (H) RNA(Chk2:GFP) and 5mis-MO-Chk2; (I) RNA(Chk1:GFP) and MO-Chk2; (J) RNA(Chk2:GFP) and MO-Chk1.

• Supplemental Figure S2 -

  Fig. S2. Alcian Blue staining of wild-type and piy mutant embryos. (A-D) Ventral (A,B) and lateral (C,D) views of wild-type sibling (A,C) and piy mutant (B,D) embryos. The piy mutant embryos display a similar defect in cartilaginous elements of head skeleton to fla mutant embryos (Plaster et al., 2006).