Fig. 8. Xfz3 is required for neural crest formation. (A) Xfz3-directed antisense morpholino oligonucleotide blocks translation of injected Xfz3 mRNA. Xfz3 mRNA was injected into one cell embryos together with Xfz3 morpholino at the doses indicated. Embryos were harvested at stage 10 and analyzed by western blot with Xfz3 antibodies. (B) Xfz3 mRNA was injected together with control (Con), Kermit (Ker) or Xfz3 morpholinos (2 ng each) or with no oligo (-) and stage 10 embryos were western blotted for Xfz3 as above. (C) RT-PCR analysis of Xslug expression (as in previous figures) in animal caps expressing chordin and Xwnt1, co-injected with either Xfz3 morpholino (FM; 4 ng) or control morpholino (CM; 4 ng) and harvested at stage 18. Xwnt1-dependent induction of Xslug was rescued by co-injection of an Xfz3 mRNA (F3) lacking the morpholino target sequence; this dose of F3 mRNA (10pg) caused minimal induction of Xslug in the absence of Xwnt1. (D-F) 2 ng of the control morpholino (D) or Xfz3 morpholino (E,F) were co-injected with mRNA for nuclear ß-galactosidase into a single dorsal-lateral (B2) blastomere at the 32-cell stage. For rescue of Xfz3 depletion, mouse Xfz3 mRNA (0.4 ng) was co-injected with Xfz3 morpholino (F). Embryos were fixed at stage 18 and ß-galactosidase activity was measured in situ (magenta) followed by whole-mount in situ hybridization for Xslug.