DEV02590 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Sequence and expression pattern of Xenopus Profilin1. (A) Clustalw alignment shows strong identity of Xenopus Profilin1 with rat, mouse and human profilin 1; ‘*’, ‘:’ and ‘.’ indicate identical, conserved and semi-conserved residues, respectively. All sequences are compared with rat. (B) Xenopus Profilin1 is expressed throughout development as monitored by RT-PCR analysis; EF1α is used as a loading control; −RT, without reverse transcriptase. (C) The spatial expression pattern of Xenopus Profilin1 is dynamic with the highest level of expression refined to the neural fold and nervous system during neurula-stage embryo. Stages and views of the embryos are shown; no signal is detected using a sense Xenopus Profilin1 probe.

• Supplemental Figure 2 -

  Fig. S2. Profilin does not interfere with canonical Wnt signaling or mesodermal specification. (A) Profilin (1 ng) does not inhibit Xwnt8 (2.5 pg)- or Dvl (500 pg)-mediated secondary axis induction. Arrowheads indicate secondary cement glands; embryos were scored at stage 30. (B) Quantitation of secondary axis induction assays, embryos with a secondary axis without or with head, eyes or cement glands were scored as partial or complete secondary axis respectively. (C) Profilin1 RNA injection (1 ng) does not affect Xwnt-8 (10 pg)- or Dvl2 (2 ng)-mediated induction of Xnr3 and Siamois in animal explants at stage 10.5, as assayed by RT-PCR. EF-1α was used as an internal control; WE, whole embryo; −RT, no reverse transcriptase added. (D) Neither overexpression nor depletion of Xenopus Profilin1 affects mesodermal gene induction by activin in animal explants, as assayed by RT-PCR at stage 10.5.