DEV01717 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Comparison of the expression domains of Otx2 and Hoxb1 on immediately adjacent sections in Gbx2 mutants at E8.5. (A,B) There is a gap between the expression domains of Otx2 (A) and Hoxb1 (B) in Gbx2^–/– embryos at the three-somite stage. (C-F) The expression domains of Otx2 and Hoxb1 partially overlap in Gbx2^–/– (C,D) and Gbx2^–/–; HG (E,F) embryos after the five-somite stages. The posterior limit of Otx2 is indicated by arrows, and Hoxb1 by arrowheads.

• Supplemental Figure 2 -

  Fig. S2. Formation of the cranial nerves analyzed by neurofilament antibody staining. (A-D) Whole-mount immunostaining of E10.5 embryos of indicated genotypes. Expression of the HG transgenes are shown by X-gal staining (blue) in B and D. Embryos were bisected for photographing. Cranial nerve IV is missing in Gbx2^–/– and Gbx2^–/–; HG embryos. Cranial nerve III and V are formed abnormally, whereas VII/VIII and X appear normal in Gbx2^–/– and Gbx2^–/–; HG embryos.

• Supplemental Figure 2 -

  Fig. S3. Abnormal expression of follistatin and Hoxb1 in Gbx2^–/– embryos. (A,B) RNA in situ hybridization of follistatin in whole-mount wild-type (A) and Gbx2^–/– embryos at the six-somite stage. The transverse bands of follistatin expression in r1-2 and r4 are missing in Gbx2^–/– embryos, whereas follistatin expression in the paraxial mesoderm (arrow) posterior to r3 is normal. Arrowhead indicates the r2/3 border, which is aligned with the preotic sulcus. (C,D) RNA in situ hybridization of Hoxb1 in whole-mount wild-type (C) and Gbx2^–/– embryos at the six-somite stage. The expression of Hoxb1 in r4 appears expanded anteriorly relative to the preotic sulcus (arrowhead) and the lateral-most expression (arrow) is missing.