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Fig. S1. Ectopic expression of the CNH marker fgf4. (A-C) Ectopic expression of the CNH marker fgf4 in the secondary tail of mfn embryo (B) and dvl2/3 morphant (C) fixed at 24 hpf. Lateral view with anterior to the left.
Fig. S2. Distribution of morphological phenotypes of flh-expressing CNHs in mfn, kny and snh embryos fixed at the indicated time points. Right panel shows the lateral view of representative embryo in each phenotypic class. For each time point around 40 embryos were scored.
Fig. S3. Notochord bifurcation in kny embryos. (A-E) Lateral view of expression of ntl (A,B) and col2a (col2α; C-E) in the tail of wild-type and kny embryos fixed at 24 hpf. Note the three separate flh-expressing CNH domains in B, triple-branched notochord (nt) in D and birfurcated notochord with detached hypochord (hc) in E. Each region is indicated by arrowheads.
Fig. S4. β-Catenin localization is unaffected in mfn and kny mutants. (A-C) Confocal microscopy images of tailbud in wild-type, kny and mfn embryos labeled with β-catenin antibody at 21 somites; inset, close-up of CNH region. Lateral view with anterior to the left.
Fig. S5. BMP signaling is active in the ventral-posterior tailbud. (A,B) Confocal microscopy images of tailbud in wild-type and mfn embryos labeled with β-catenin (red) and P-Smad5 (green) antibody at 21 somites. Note that in wild-type embryo (A), P-Smad5 staining is expressed in the posterior tailbud and extends under the notochord, whereas it is absent in mfn (B).
Movie 1. Time-lapse video of normal tailbud morphogenesis in an mRFP-labeled live Tg(flh:EGFP) embryo starting at the 10-somite stage. Lateral view, anterior to the left.
Movie 2. Time-lapse video of defective tailbud morphogenesis in an mRFP-labeled live mfn;Tg(flh:EGFP) embryo starting at the 10-somite stage. Lateral view, anterior to the left.
Movie 3. Time-lapse video of defective CNH morphogenesis in an mRFP-labeled aive kny;Tg(flh:EGFP) embryo starting at the 10-somite stage. Lateral view, anterior to the left.