Fig. 1

Hipk2 Is Required for Dvl-Mediated Early Embryonic Events in Zebrafish

(A) Hipk2 is required for the stabilization of β-catenin in zebrafish. Zebrafish embryos were injected with MOs. Extracts were harvested from embryos at 8 hr postfertilization (hpf) and immunoblotted with anti-zebrafish β-catenin. α-Tubulin was used as a loading control.

(B and C) Hipk2 is essential for the β-catenin pathway-mediated posterior mesoderm formation and convergent extension (CE). Embryos were injected with MOs. Panels in (B) show whole-mount in situ hybridization of d2EGFP, tbx6, or ntla in OTM:d2EGFP-transgenic zebrafish embryos. Panels in (C) show whole-mount in situ hybridization of ntla and dlx3b or myod1 in zebrafish embryos. Blue, red, and orange two-way arrows indicate the width of neural plate, the length of notochord, and the width of myod1-expression domain, respectively. The percentages of embryos showing similar expression patterns and total number of MO-injected embryos (n) are shown under each image. In all images, A, P, V, and D indicate anterior, posterior, ventral, and dorsal sides. Scale bar, 200 μm.

(D and E) Hipk2 is required for the protein stability of endogenous Dvl (D) and exogenous mouse Dvl1 (E) Dvl in zebrafish. Zebrafish embryos were injected with MOs without (D) or with mouse Myc-Dvl1 and hemagglutinin (HA)-GFP mRNA (E). Extracts were harvested from the embryos at 8 hpf. In (D), embryo extracts were immunoprecipitated and then immunoblotted with anti-Dvl (see te Experimental Procedures for details). In (E), extracts were immunoblotted with indicated antibodies. GFP was used as a loading control.

See also Figures S1 and S2.