Xenbase Image ID: 80384

FIG. 3. Inhibition of Xenopus Wnt-4 translation in vitro by Wnt-4 antisense morpholinos. Plasmids (50 ng) encoding Wnt-4, HAWnt-4, and Wnt-1 were used as templates in cell-free coupled transcriptionranslation reactions. Several morpholinos [Wnt4-MO; sense control, Wnt4(sc)-MO; four-mispaired control, Wnt4(mp)-MO; and a standard control, Cont-MO] were tested for inhibition of translation. Cell-free transcriptionranslation reactions were performed in the presence of [35S]methionine and analyzed by SDSAGE/autoradiography. (A) Dose-response analysis of inhibition of Wnt-4 translation by Wnt4-MO. The amount of radiolabeled protein produced was quantified by using a phosphorimager. The control sample, which was not treated with Wnt4-MO, was set at 100%. Significant reduction of protein synthesis occurred with 1 and 3 􏰂M Wnt4-MO. (B) Sequence-specific inhibition of Wnt-4 translation by Wnt4-MO. Morpholinos (1 􏰂M) were used to inhibit translation of Wnt-4. Only Wnt4-MO was effective in blocking Wnt4 translation. (C, D) In vitro translation of HAWnt4 (an HA-tagged Wnt-4 variant lacking the complementary target sequence for Wnt4-MO) and Xenopus Wnt-1 was essentially unperturbed in the presence of Wnt4-MO. Morpholinos were used at 1 uM. Image published in: Saulnier DM et al. (2002) Copyright © 2002. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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