Figure 1.

Removal of tritin from embryos. Extracts were prepared from unwashed or washed embryos (A), and the depurination assay was performed (B). Translation mixtures prepared with the extract from unwashed embryos were incubated for 0, 1, 2, 3, and 4 h (B, lanes 1–5 respectively); mixtures with washed embryos were incubated for 0, 2, and 4 h (lanes 10–12, respectively). Isolated RNA was treated with acid/aniline, then was separated on 4.5% polyacrylamide gels. Additionally, RNA was directly extracted from embryos with guanidine isothiocyanate-phenol and was analyzed as above before (B, lane7) and after (B, lane 8) treatment with acid/aniline. For the fragment marker (B, lanes 6 and 9), incubation was carried out in the presence of gypsophilin (40), a highly active RIP from Gypsophila elegance; the arrow indicates the aniline-induced fragment.