DEV01554 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. The amphioxus retinoic acid receptor (AmphiRAR) is autoregulated. In control embryos treated with DMSO, the highest level of AmphiRAR expression in the endoderm is in the middle third of the gut at 16 hours (A), 20 hours (B) and 24 hours (C). In amphioxus embryos treated with 1310^-6 M RA during the gastrula stage, expression of AmphiRAR is upregulated and shifted forwards. The anterior limit of AmphiRAR expression is at the anterior end of the pharynx at 16 hours (D), 18 hours (E) and 24 hours (F). In amphioxus embryos treated with 1310^-6 M BMS009 at the gastrula stage, AmphiRAR is downregulated and expression shifted posteriorly. The anterior limit of expression is not substantially different from controls at 16 hours (G), but is shifted increasingly posterior at 20 hours (H) and 24 hours (I) to coincide with the posterior limit of the expanded pharynx. Arrowheads indicate anterior limit of AmphiRAR expression in the pharynx. Scale bars: 50 mm.

• Supplemental Figure 2 -

  Fig. S2. An antisense morpholino oligonucleotide complementary to AmphiHox1 suppresses translation in an in vitro translation assay. AmphiHox1 protein is about 40 kDa. The negative control (neg. cont.) does not include either morpholino oligonucleotide or AmphiHox1expression plasmid. All other lanes include 200 ng AmphiHox1 expression plasmid DNA. At 100 ng the AmphiHox1 morpholino almost completely blocks translation of AmphiHox1 mRNA and at 500, 1000 and 5000 ng of AmphiHox1 morpholino, translation of AmphiHox1 mRNA is completely blocked. The same concentrations of control morpholino (right four lanes) have no effect on translation of AmphiHox1 mRNA.