DEV01707 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Nucleotide sequences of the regulatory modules of Otx responsible for driving its lineage specific transcription in the two ascidian species, Halocynthia roretzi (Hr) and Ciona intestinalis (Ci). The regulatory modules for the A-line (A), bB-line (B) and ab-line cells (C) are shown. Putative TF binding sites (BSs) for Lhx, Fox, T-box, GATA and Ets are indicated by red, yellow, blue, green and yellow-green boxes. They were searched by the computer programs and by referring to the results of previous reports (for details, see Materials and methods). The open red box in the module Hr #6 indicates the Lhx BS, on which we omitted mutational analysis shown in Fig. 4, because this BS exhibited no activity when the other three Lhx BSs in Hr #6 were mutated.

• Supplemental Figure 2 -

  Fig. S2. Comparison of Otx upstream sequences between Halocynthia roretzi and Ciona intestinalis. The nucleotide sequences, spanning upstream down to the sixth coding exon, were compared between Hr-Otx and Ci-Otx using PipMaker programme (http://bio.cse.psu.edu/pipmaker/pip-instr.html). Significant similarity is indicated by horizontal (D) or diagonal (E) segments. (D) The nucleotide sequences of Hr-Otx (9963 nucleotide long) is on the x-axis, and the % of sequence identity with Ci-Otx is represented in the y-axis. The parameters used in this analysis are the same as those used in the previous work by Kimura-Yoshida et al., in which Otx2 transcriptional regulatory elements have been successfully identified from the sequence comparison between Fugu and mouse (Kimura-Yoshida et al., 2004). (E) The nucleotide sequences of Hr-Otx (9963 nucleotides) and Ci-Otx (7754 nucleotides) are depicted in the x-axis and y-axis, respectively.