S1Supplemental Data Subcellular Localization and Signaling Properties of Dishevelled in Developing Vertebrate Embryos

Experimental Procedures 10. DMZs were cut with an eyebrow knife, sharpened watchmak-er’s forceps (Inox #5), and immobilized for imaging as described for animal caps above.Embryological Methods Female adult Xenopus laevis were ovulated by injection of human Analysis of Convergent Extension in Whole Embryoschorionic gonadotropin, and eggs were fertilized in vitro and dejel-Embryos were injected with indicated mRNAs into the two dorsallied in 3 % cysteine (pH 7.9) and subsequently reared in 1/3 MMR blastomeres at the 4-cell stage. Embryos were then reinjected with(Marc’s modified ringers). For microinjections, embryos were placed fluorescent dextran into one dorsal blastomere at the 256-cell stage.in a solution of 2.5 % Ficoll in 1/3 MMR and injected via forceps These embryos were then cultured until the onset of gastrulation,and an Oxford universal micromanipulator; they were then reared and embryos were photographed for florescent and bright-field im-in 3 % ficoll 1/3 MMR to stage 9 and then washed and reared ages with a Leica MZ16FA stereomicroscope. The embryos werein 1/3 MMR alone. Embryos were staged according to [S1]. then reared to late gastrula stages and rephotographed.Embryo culture, solutions, in vitro transcription of mRNAs, in situ These embryos were then fixed overnight in Bouin’s fixative,hybridization, and RT-PCR were performed as previously described washed, bleached, dehydrated, and cleared for confocal imaging.[S2–S4]. Fixed embryos were bisected for ease of positioning for imaging