Fig. S6

Zebrafish Dishevelled Experiments. A, While injection of the various Dvl and Daam1 RNAs generated predicted defects in convergence and extension (CE) movements (red bars), there was no effect on the caudal migration of FBM neurons (green bars). Injection of GFP RNA did not affect CE movements or neuronal migration. Data from 2-3 experiments; number in parenthesis denotes number of embryos. B, Western blot analysis of proteins isolated from 20 hpf embryos injected with RNAs encoding various constructs. Myc-tagged proteins of the predicted sizes were detected. A Western for α-tubulin was performed simultaneously to detect tubulin as a loading control. A separate Western blot was performed to detect Flag-tagged N-Daam1. C, Dorsal views of hindbrains processed for whole-mount immunostaining with anti-GFP (green) and anti-MYC (red) antibodies. 20 hpf control uninjected embryos(left panels) and embryos injected with DvldelPDZ RNA (right panels). The mutant Dvl protein is expressed broadly throughout the hindbrain and in the FBM neurons (yellow cells, arrowheads), albeit in mosaic fashion. FBM neuron migration is not affected despite extensive expression of mutant protein both in the neurons and the surrounding tissue. The anti-MYC antibody exhibits weak non-specific labeling in control embryos (asterisks).