DEV020784 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Phylogenetic analysis of Nematostella FGF ligands. Consensus tree of phylogenetic analyses of FGF domains performed by Bayesian inference using MrBayes3.1 (Ronquist and Huelsenbeck, 2003; Huelsenbeck and Ronquist, 2001), based on a ClustalW alignment (Thompson et al., 1994) of Nematostella ligands (Nv) identified and cloned in this paper, and the FGF ligands of humans (Hs). A mixed model of substitution was used, four gamma rate categories, convergence of trees was reached after 1 Mio generations (s.d.<0.008), and a burnin of 2800 trees. Posterior probability values are given above the given nodes. Bar represents substitutions per site. NvFGFA1 and NvFGFA2 are the two paralogs analysed in this paper.

  Huelsenbeck, J. P. and Ronquist, F. (2001). MRBAYES: Bayesian inference of phylogenetic trees. Bioinformatics 17, 754-755.

  Ronquist, F. and Huelsenbeck, J. P. (2003). MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19, 1572-1574.

  Thompson, J. D., Higgins, D. G. and Gibson, T. J. (1994). CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22, 4673-4680.

• Supplemental Figure S2 -

  Fig. S2. Phylogenetic analysis of FGF receptors. Consensus tree of phylogenetic analyses of FGF receptors of the split tyrosine kinase domains performed by the Maximum likelihood programs TreePuzzle (Schmidt et al., 2002) and PhyML (Guindon and Gascuel, 2003), as well as by Bayesian inference using MrBayes3.1 (Ronquist and Huelsenbeck, 2003; Huelsenbeck and Ronquist, 2001), based on a ClustalW alignment (Thompson et al., 1994) of Nematostella FGFR (Nv) identified and cloned in this paper, and the FGF receptors of humans (Hs). Nv, Nematostella vectensis; Bb, Branchiostoma belcheri; Hs, Homo sapiens; Xl, Xenopus laevis; Bm, Bombyx mori; Hv, Hydra vulgaris; Dm, Drosophila melanogaster; Sf, Spodoptera frugiperda; Ci, Ciona intestinalis; Hr, Halocynthia roretzi; Pc, Podocoryne carnea; FGFR, fibroblast growth factor receptor; VEGFR, vascular endothelial growth factor receptor; MSRTK, muscle skeletal receptor tyrosine kinase.

  Guindon, S. and Gascuel, O. (2003). A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Syst. Biol. 52, 696-704.

  Schmidt, H. A., Strimmer, K., Vingron, M. and von Haeseler, A. (2002). TREE-PUZZLE: maximum likelihood phylogenetic analysis using quartets and parallel computing. Bioinformatics 18, 502-504.

• Supplemental Figure S3 -

  Fig. S3. Protein domain structure of the Nematostella FGF receptors. (A) Nematostella FGFRa. (B) Nematostella FGFRb. Protein structure analysis according to the ‘SMART’ database at EMBL (http://smart.embl-heidelberg.de/). Red indicates signal peptide; green, IG-loops; dark blue bar, transmembrane domain; blue box, split tyrosine kinase domain.

• Supplemental Figure S4 -

  Fig. S4. Absence of the apical tuft upon NvFGFRa MO injection. Scanning electron micrograph of a 4-dpf larvae injected with the NvFGFRa MO. Asterisk indicates the aboral pole.

• Movie 1 -

  Movie 1. Swimming behaviour of 4-day-old planula larvae injected with control MO.

• Movie 2 -

  Movie 2. Individual control MO-injected larvae swimming directionally, with the aboral pole forward.

• Movie 3 -

  Movie 3. Swimming behaviour of 4-day-old planula larvae injected with the NvFGFa1 MO. All larvae lack the apical tuft.

• Movie 4 -

  Movie 4. Individual NvFGFa1 MO-injected larvae swimming directionally, with the aboral pole forward despite lack of the apical tuft.

• Movie 5 -

  Movie 5. Swimming behaviour of 4-day-old planula larvae injected with the NvFGFa2 MO. All larvae have an expanded apical tuft.