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Fig. 1
The Mid domain of TNRC6A is sufficient to induce translational repression and deadenylation. (A) Schematic structures of zebrafish TNRC6A and its deletion mutants. (B) Schematic representation of the λN tethering assay in zebrafish embryos. (C) Results of the tethering assay with TNRC6A fragments. The bar graph shows Rluc activity that was normalized to Fluc activity. The normalized Rluc activity with the HA-λN empty construct (HA-N) was set to one. The data show averages of three independent experiments. Error bars show SD. Asterisks indicate p < 0.01 compared to experiments with HA-N. (D) The qRT-PCR analysis of reporter mRNA stability. The normalized Rluc mRNA values [normalized to those of the HA-λN empty construct (HA-N)] were set to one. The data show averages of three independent experiments. Error bars show SD. (E) The poly(A) tail analysis of the Rluc-BoxB-pA reporter mRNA by RNaseH digestion and Northern blot. The lane +dT shows completely deadenylated fragments, which correspond to A0. (F) Western blotting detecting HA-tagged effecter proteins.