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Files in this Data Supplement:
Fig. S1. Morphological defects of chato mutant embryos. (A,B) Scanning electron micrographs of wild-type (A) and chato mutant (B) mouse embryos at e8.5. chato mutants grow outside of the yolk sac (ys), which showed bubble-like protrusions in most embryos as early as e8.5 (B). (C-F) Whole-mount in situ hybridization on e7.5 embryos using markers for the primitive streak (T; C,D) and anterior definitive endoderm (Cer1; E,F). Arrowheads point to the yolk sac endoderm, which had a smooth surface in chato mutants at this stage. At e7.5, chato mutants showed an abnormally shaped epiblast (D,F; dashed lines), as compared with the proximo-distally elongated egg cylinder of wild-type littermates (C,E). Scale bars: 200 µm.
Fig. S2. Lack of genetic interaction between chato and Lp. (A-C,E-G) In situ hybridization with Meox1 and T probes was used to evaluate the mesoderm and notochord phenotypes of embryos of the following genotypes: (A) wild type, (B) Lp−/−, (C) chato−/−, (E) Lp−/−; chato+/−, (F) Lp+/−; chato−/− and (G) Lp−/−; chato−/−. The numbers in brackets indicate the number of embryos analyzed for every mutant combination. (D) Lp+/−; chato+/− double heterozygote animals were viable and fertile and had the characteristic curly tail of Lp+/− heterozygotes.
Fig. S3. Chato does not regulate transcription of non-canonical Wnt signaling components; nor does non-canonical Wnt signaling regulate chato expression. (A-H) Ventral views of wild-type (left column) and chato mutant (right column) embryos hybridized with probes for Vangl2 (A,B), Celsr1 (C,D), Fzd3 (E,F) or Dvl2 (G,H). Expression of Vangl2, Celsr1 and Fzd3 is limited to the neural tube (arrowheads), which was open and convoluted in chato mutants (B,D,F). Dvl2 was expressed in neural tube (arrowhead in G) and mesenchymal tissues (arrow in G) of both wild-type (G) and chato mutant (H) embryos. (I,J) Lateral views of wild-type (I) and Lp mutant (J) embryos hybridized with a chato probe.
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