FIG. 7.

ff1bMO and prox1MO inhibited protein translation in zebra fish embryos. (A) The pLuc3xFRE luciferase reporter and the expression plasmid, pcDNA3.1-ff1b were coinjected into one-cell-stage embryos, with or without ff1bMO (1 pmol per embryo). pRL-CMV was included as a normalization control. pcDNA3.1 was also coinjected with the luciferase reporter to determine the background value. Embryos (n = 5) were harvested at 14 hpf, and dual luciferase assays were performed. The normalized luciferase activity was calculated from three experiments. (B) prox1MO (1 pmol per embryo) was injected into one-cell-stage embryos. Both uninjected and injected embryos were harvested at 31 hpf for protein extraction. An aliquot (equivalent to 30 embryos) was separated by SDS-PAGE and analyzed by Western blotting with anti-prox1 antibody. An 83.2-kDa band, which is equivalent to the putative molecular mass of Prox1, was knocked down by prox1MO, while two nonspecific high-molecular-mass bands remained unaffected.