DEV01528 Supplementary Material

Files in this Data Supplement:

• Supplemental Data 1 (Adobe PDF) -

  Data S1. Effects of immuno-neutralising EDEN-BP on EDEN-mediated deadenylation

• Supplemental Figure 1 -

  Fig. S1. Immuno-neutralisation of EDEN-BP in Xenopus embryos specifically inhibits EDEN-mediated deadenylation. (A) Total protein extracts (equivalent to one egg/embryo) from unfertilized eggs, and from stage 3, 21 and 33/34 embryos, as indicated, were resolved by 10% SDS-PAGE, blotted onto nitrocellulose and probed with either non-immune (lanes 1-4) or anti-EDEN-BP (lanes 5-8) purified antibodies at a concentration of 5 ng/ml. Positions of the molecular weight markers are indicated on the left. (B) Radiolabeled, capped, poly(A)⁺ EDEN-containing transcript (GbEg2-410, 1 fmole) was injected together with 200 ng of non-immune (lane 1-5) or anti-EDEN-BP (lane 6-10) antibodies in Xenopus two-cell embryos. RNA was extracted at the indicated times after injection, resolved on a polyacrylamide gel containing urea and revealed by autoradiography. The position of the fully adenylated (A65) and deadenylated forms of the transcript are indicated. (C) Quantification of the experiment shown in B. The percentage of deadenylated transcript is plotted against time. (D) Same as is shown in B, with an initially deadenylated transcript. (E) Quantification of the experiment shown D, quantified as in C.