DEV02835 Supplementary Material

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• Supplemental Figure 1 (Adobe Acrobat) -

  Fig. S1. DjCHC encodes a homolog of CHC with highly conserved domains. (A) Deduced amino acid sequence of DjCHC. DjCHC has a globular N-terminal domain (red), knee region (yellow), distal domain (green), proximal domain (blue) and C-terminal end (gray). Clathrin heavy-chain repeat homology regions are indicated by boxes. Each of these repeat regions is approximately 140 amino acids long and the regions are composed of multiple alpha helical repeats in the arm region of the clathrin heavy chain. The complete cDNA sequence of DjCHC has been deposited in DDBJ/EMBL/GenBank databases with the accession number AB243057. (B) Phylogenetic tree produced by the neighbor-joining method using full-length CHC sequences. The tree includes amino acid sequences derived from Dugesia japonica (AB243057), Homo sapiens (Q00610), Mus musculus (Q68FD5), Rattus norvegicus (P11442), Bos taurus (P49951), Gallus gallus (CAD20886), Xenopus laevis (AAH73439), Danio rerio (AAT68095), Drosophila melanogaster (P29742), Caenorhabditis elegans (P34574), Glycine max (AAC49294), Arabidopsis thaliana (AAF01510) and Saccharomyces cerevisiae (P22137).

• Supplemental Figure 2 (Adobe Acrobat) -

  Fig. S2. Representative survival curve of regenerates and uncut planarians, and phototactic behavior analysis of DjCHC-RNAi-treated planarians. (A) Fractions at various times after amputation are represented by black (control) and red (DjCHC-RNAi-treated planarian) lines. Head pieces, which contained many neural cells, of DjCHC-RNAi-treated planarians died at a high rate from day 8 to day 13 after amputation compared with the other fragments. (B) Fractions of planarians treated with dsRNA of DjCHC or X-ray irradiation. X-ray-irradiated planarians died approximately 2 weeks after irradiation. By contrast, almost all DjCHC-RNAi-treated planarians survived for more than 3 weeks, although their heads were atrophied. (C) The assay consists of tracing the movements of planarians in response to light as previously described (Inoue et al., 2004). Each colored line indicates the trajectory of an individual planarian during 90 seconds. Trajectories representing the movements of control and DjCHC-RNAi planarians 7 days after regeneration (n=10) are shown. Tracing of the movements of control animals revealed that all of them moved away from the light source and reached the dark side. By contrast, DjCHC-RNAi-treated planarians could not recognize the direction of the light and they hardly moved.