DEV029025 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Dkk-1 depletion affects Xnr3 expression pattern. Whole-mount in situ hybridization for Xnr3 mRNA in uninjected and Dkk1-depleted early gastrulae (stage 10). The Xnr3 expression territory is narrowed in the left-right axis and expanded along animal-vegetal axis. The dorsal lip is marked with a white arrowhead.

• Supplemental Figure S2 -

  Fig. S2. Dkk-1 depletion increases Wnt signaling dorsally and ventrally. TOPflash reporter was injected into both dorsal and ventral blastomeres (four-cell stage) and embryos were dissected at stage 9. Both dorsal and ventral halves show increased luciferase activity in Dkk1-depleted embryos compared to uninjected controls.

• Supplemental Figure S3 -

  Fig. S3. Dkk1 depletion induces ectopic Wnt target gene expression. Real-time RT-PCR shows induced Xnr5 expression within ventral marginal zone as well as dorsal marginal zones of Dkk1-depleted embryos compared with controls.

• Supplemental Figure S4 -

  Fig. S4. Dkk1 depletion induces elongation movement in both dorsal and ventral explant. For dorsal and ventral explants, the embryos were marked on the dorsal side at the four-cell stage with Nile Blue, and the equatorial zone was dissected and cut into dorsal and ventral halves at the late blastula stage.

• Supplemental Figure S5 -

  Fig. S5. The Dkk1 depletion phenotype depends on both Wnt5a and Wnt11. (A) The phenotypes of control, Wnt5a-depleted, Dkk1-depleted and Wnt5a/Dkk1-depleted embryos at the neurula stage. Wnt5a/Dkk1-depleted embryos have the same ventralized phenotype as Wnt5a-depleted embryos. (B) The phenotypes of control, Wnt11-depleted, Dkk1-depleted and Wnt11/Dkk1-depleted embryos at the mid-gastrula stage. Wnt11/Dkk1-depleted embryos have the same phenotype as Wnt11-depleted embryos.