DEV009464 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Lack of Cryptic expression in the endoderm. (A-D) In situ hybridization of transverse frozen sections of E8.2 embryos with probes specific for Nodal (A), Cryptic (B), ActRIIB (C) or Alk4 (D). (E-H) Higher-magnification views of the images in A-D, respectively. All sections were counterstained with nuclear fast red. The expression of Nodal and Cryptic is absent from the endoderm, whereas that of ActRIIB and Alk4 is present there. Although the expression of them in the endoderm is difficult to be judged because of less cytoplasm with larger nucleus in the squamous-shaped endoderm, the judgement should be enabled by assessing cytoplasmic staining near the cell-cell junction of the endoderm (arrowheads). ec, ectoderm; en, endoderm; mes, mesoderm. Scale bars: 100 μm.

• Supplemental Figure S2 -

  Fig. S2. Lack of Cripto expression in or around the node. (A, B) Whole-mount in situ hybridization with a probe specific for Cripto in E8.2 embryo. Anterior (A) and left (B) views are shown. Notice that Cripto is expressed in the prospective heart region at the early somite stages (black arrowhead), but is not in the node (red arrowhead) or other tissues. Scale bar: 500 μm.