DEV02413 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Detectable level of Furin is restricted to the PrE lineage. Paraffin section (7 μm) of an agar pre-embedded E4.5 embryo previously processed through in situ hybridization for Furin mRNA. The staining is detected in a layer of cell covering the epiblast and facing the blastocoelic cavity, thus corresponding to the PrE. Some staining is also apparent in cells lining the trophectoderm, probably corresponding to parietal endoderm cells.

• Supplemental Figure 2 -

  Fig. S2. ExVE markers are downregulated in EmVE between E5.25 and E5.5. Embryos between E4.5 and E5.5 were analysed by in situ hybridization using Gata4, Hnf4 and Ttr antisense RNA probes. Whereas the three markers are restricted to the PrE at E4.5 and show ubiquitous expression in the VE at E5.25, they become downregulated by E5.5 specifically in the EmVE.

• Supplemental Figure 3 -

  Fig. S3. Fgf5 and Fgf8, but not Lim1 show mRNA expression in epiblast in addition to EmVE. Paraffin section (7 μm) of agar pre-embedded E5.25 wild-type and Nodal^lacZ/lacZ null mutant embryos stained for Fgf5, Fgf8 or Lim1 mRNAs. Whereas staining is well apparent both in epiblast and EmVE for Fgf5 and Fgf8 (only one embryo shown here as they have identical profiles), it is apparent only in EmVE for Lim1. The three markers are absent from the VE of Nodal^lacZ/lacZ null mutant. Fgf5 and Fgf8 are also strongly downregulated in epiblast of Nodal^lacZ/lacZ null mutant.