Fig. 5

miR-1 represses target mRNA in a deadenylation- and PABP-independent manner in zebrafish embryos. (A) Fluc reporter mRNA containing zebrafish pdlim1 32UTR. Red boxes indicate the target site for miR-1. (B) Scheme of the miR-1 repression assay in zebrafish embryos. (C) The poly(A) tail analysis of the Fluc-pdlim1 32UTR reporter mRNAs at six hours in the absence (-) or presence (+) of the miR-1 duplex. The left panel shows the reporter mRNA with a normal poly(A) tail [Fluc-pdlim1-poly(A)]. The right panel shows the reporter mRNA with the A98C10 tail (Fluc-pdlim1-A98C10). (D and E) The results of the miR-1 repression assay with reporter mRNA containing a normal poly(A) tail in the presence of control Myc-GFP. (F and G) Results of the miR-1 repression assay with reporter mRNA containing the A98C10 tail in the presence of control Myc-GFP. (H and I) Results of the miR-1 repression assay with reporter mRNA containing the A98C10 tail in the presence of Myc-Paip2. D, F, and H show normalized Fluc activity. E, G, and I show normalized Fluc mRNA levels, which were measured using qRT-PCR. The values of the experiments using miR-124 were set to one at each time point. The data shows averages of three independent experiments. Error bars show SD.