DEV02552 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Delayed recruitment of Pxm. Lim1 is expressed in the node (lower arrowhead) and notochord at the allantoic bud stage (A). In mutant embryos, node was formed more proximally (lower arrowhead, A′). The primitive steak was shorter in the mutant compared with control embryos (between arrowheads, A and A′). At the head-fold stage, Foxa2 was expressed in the node (lower arrowhead, B and B′) and notochord. The length of the primitive streak (between arrowheads) was similar between control and mutant embryos (B,B′). Tbx6 was expressed in the mesoderm except node. Tbx6 expression domain was shifted to proximal and decreased in mutant embryos (between arrowheads, C and C′). Scale bars: 250 μm for A-B′; 100 μm for C,C′.

• Supplemental Figure 2 -

  Fig. S2. Overexposure to SU5402 inhibits normal development. (A) All embryos cultured with 40 μM SU5402 showed defective development (arrowheads). (B-E) The development of somites was inhibited in control and mutant embryos cultured with 20 μM SU5402 for around 48 hours (B,C, white arrow). Uncx4.1 was either not expressed or expressed faintly in the epiblast in such embryos (D,E). Scale bars: 100 μm for A; 50 μm for B,C; 600 μm for D,E.

• Supplemental Figure 3 -

  Fig. S3. Partial rescue of somite development in mutant embryos. (A,B) Mutant embryos cultured with DMSO for around 48 hours showed an expansion of somites (A, arrow is neural tube and arrowhead is lateral edge of the embryo; B, arrowheads). (C,D) Mutant embryos were cultured with 20 μM SU5402 for 12-14 hours, washed three times and further cultured with DMSO for around 48 hours in total. In three mutant embryos out of ten tested, the rescue was observed on one side of the embryos but not on the other side. Two are shown. Two columns of somites were observed (arrowhead) on one side of the embryo. Arrow is neural tube (C). Rescue was observed in more rostral somites (arrowhead) but not in more caudal somites (arrow), indicating that abnormal recruitment of prospective Pxm occurred after removal of SU5402 (D). All embryos were subjected to in situ detection of Uncx4.1 after 48 hours in culture. Scale bars: 600 μm.