C-Daam1 and C-Daam1 FH2 mutant domain structures are shown on the top. A, B, D, E, in situ hybridization of stage 15/16 embryos with indicated antisense RNA probes. Anterior views are shown. Light blue is the lineage tracer (nĪ²Gal). Arrowheads point to expanded foxd3 and sox8 in C-Daam1 RNA-injected embryos (10-20 pg, A, B). C, C-Daam1 upregulates NC gene expression in neuralized ectoderm. Ectoderm explants were prepared from embryos injected with C-Daam1 RNA (10-20 pg) or wnt8 RNA (0.5 ng) and chordin RNA (150 pg) as indicated, cutured to stage 16 and processed for RT-PCR analysis. The explants coexpressing chordin and C-Daam1 upregulate pax-3 and sox8 but do not activate myod. D, E, C-Daam1-FH2* RNA (10-20 pg) does not promote NC gene expression. F, Frequencies of embryos with altered NC marker expression. Total number of explants is shown on top of each bar.