DEV02613 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Injection of a splice-blocking morpholino results in the loss of lef1 mRNA transcripts. (A) Diagram of lef1 gene structure indicating the location of the lef1 MO target at the exon 7 splice donor site. Introns are not to scale. (B) RT-PCR analysis demonstrates that injection of the lef1 MO into zebrafish embryos at the one-cell stage results in the production of aberrantly spliced forms at 12 hpf and the loss of lef1 mRNA after 18 hpf.

• Supplemental Figure 2 -

  Fig. S2. Embryos homozygous for the X8 deletion phenocopy Lef1 morphants. (A-C) nk2.1a, rx3 and emx2 expression are unaffected in X8 mutants at 30 hpf. (D-G) sox3, zash1a, dlx2 and isl1 are all absent in the posterior hypothalamus of X8 mutants at 30 hpf. White circles outline the posterior hypothalamus.

• Supplemental Figure 3 -

  Fig. S3. Rescue of lef1 MO phenotypes by lef1 and sox3 mRNA injection. Co-injection of 2 ng lef1 MO with 100 pg lef1 mRNA (A,C,E) or 20 pg sox3 mRNA (B,D,F) rescued the expression of zash1a (A,B), dlx2 (C,D), and isl1 (E,F) in the posterior hypothalamus at 30 hpf. White circles outline the posterior hypothalamus.