Figure 5.

Ryk is required for Dsh to regulate the activation of noncanonical Wnt signaling. (A–G) Rescue assays between Ryk and noncanonical Wnt components. Two blastomeres of four-cell stage embryos were injected at the dorsal marginal region with the indicated reagents. Quantitative results of DMZ elongation assay are explained in Table S1 (available at http://www.jcb.org/cgi/content/full/jcb.200710188/DC1). (H) Quantitative assays of rescue between Ryk and Ryk ΔC in DMZ elongation assay. n, total number of explants. Error bars indicate the mean ± SD. (I) Ryk ΔC, unlike Ryk, did not phosphorylate Dsh. Animal caps expressing myc Dsh (1 ng) alone or with the combinations of XRyk (1 ng) and XRyk ΔC (1 ng), as indicated, were subjected to Western blotting using anti-myc. Numbers to the left of the gel blot indicate molecular mass standards in kD. (J–M) The normal RhoA distribution was not translocated by Ryk ΔC and Ryk MO to the cell membrane in animal cap cells. XRyk (1 ng), XRyk MO (80 ng), or XRyk ΔC (1 ng) were injected with GFP RhoA (500 pg) into the animal regions of embryos at the four-cell stage, either alone or in combination as indicated. Bar, 20 μm. (N) Ryk and Fz7 have a cooperative effect in CE movements. Others indicate a truncated and mild kinked axis. n, total number of embryos.