DEV02458 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. In vitro transcription/translation of xWT1, XHRT1, esr9, esr10 and Xhairy2b is specifically inhibited by the respective MOs. (A) The xWT1 MOs inhibit xWT1a translation but have no effect on the translation of GFP. A standard control, Cont-MO, has no effect on xWT1. (B) The XHRT1-MO inhibits the translation of both XHRT1 pseudoalleles but has no effect on XHRT1a-mut (with six mismatches in the 5′ UTR MO target sequence) and esr9. The Cont-MO has no effect on XHRT1a. (C) The esr9 and esr10 MOs both inhibit the translation of esr9a and esr10a, but have no effect on esr8. The Cont-MO has no effect on esr9a and esr10a. (D) The Xhairy2b MOs inhibit the translation of both Xhairy2 pseudoalleles but have no effect on XHRT1a. The Cont-MO has no effect on Xhairy2a and Xhairy2b. In vitro transcription-translation reactions were performed by using the Sp6 Rabbit Reticulocyte Lysate System (Promega) in the presence of ³⁵S methionine. Translation products were analysed by SDS-PAGE/autoradiography.

• Supplemental Figure 2 -

  Fig. S2. Injection of esr9, esr10 and Xhairy2b MO does not inhibit xWT1 and XSMP-30 expression. (A-L) Whole-mount in situ hybridization of embryos injected with 15 ng of the indicated MOs together with β-galactosidase mRNA analysed at late tailbud/early tadpole stage with a xWT1 or XSMP-30 probe. Control and injected sides are shown. The injected area is revealed by X-gal staining. Unaffected: 75%, n=38; 77%, n=35; 63%, n=27; for esr9, esr10 and Xhairy2b MOs on xWT1; 68%, n=44; 78%, n=50; 75%, n=24; for esr9, esr10 and Xhairy2b MOs on XSMP-30.