Fig. S2

Neural crest sublineage specification fails to occur in foxd3^zdf10-tfap2aMO embryos. (A-H) Lateral views with anterior to the left. (A) mitf expression at 24 hpf (arrowheads). Just as with foxd3^zdf10;tfap2a^low double mutant embryos, foxd3^zdf10-tfap2aMO embryos completely lack neural crest (NC) mitf expression (asterisks). (B) xanthine dehydrogenase (xdh), diagnostic of xanthophore precursors, at 24 hpf (arrowheads); (C) endothelin receptor B (ednrb1), presumably expressed by all chromatophore precursors, at 30 hpf (arrowheads). NC expression of both genes is absent in mutant morphants (asterisks). (D) dlx2 expression at 24 hpf. dlx2 expression (arrowheads) is absent in the branchial arches of foxd3^zdf10-tfap2aMO embryos (asterisks), but is retained in the forebrain. (E) dlx3 expression at 30 hpf (arrowheads). (F) dhand expression at 30 hpf (arrowheads). NC expression of both genes is absent in mutant morphants (arrow; double asterisks) with dlx3 expression maintained in non-NC-derived otic vesicle (asterisks). (G) tbx1 expression in the branchial arches at 27 hpf is diagnostic of the endodermal component of the arches. tbx1 expression is normal in all experimental embryos. (H) endothelin 1 (edn1) expression in the mesodermal component of the branchial arches at 30 hpf. Branchial mesoderm appears to be develop normally in foxd3^zdf10 single mutant, tfap2aMO morphant and foxd3^zdf10-tfap2aMO mutant-morphant embryos.