Fig. 8. Tethered TNGW1 and GW182 exerted translational repression to a greater extent than Ago2. (A) Tethered TNGW1 and GW182 both exerted stronger repression effect than tethered Ago2. Assays for both luciferase activities were performed 48 hours after transfection. Tethered TNGW1 and GW182 showed 67.6% and 65.3% translation-repression effect to FL-5BoxB reporter, which was significantly higher than tethered Ago2 (^*P<0.05, t-test). Measured FL-5BoxB activities were normalized to corresponding RL activities. All translation-repression effects were estimated by the differences of FL-5BoxB/RL activity compared with that in NHA-vector-transfected group. Error bars represent standard deviations. (B) Tethered TNGW1 or GW182 induced less reporter-mRNA-reduction than tethered Ago2. The mRNA levels from the same assay were measured by SYBR-Green quantitative RT-PCR. Degradation of FL-5BoxB mRNA were determined by the reductions of the mRNA level between experimental groups and NHA-vector transfected group. All FL-5BoxB mRNA levels were normalized to Renilla luciferase mRNA to minimize the experimental errors. (C) Tethered TNGW1 and GW182 strongly reduced the translation efficiency of the reporter compared with tethered Ago2. Translation efficiencies of FL-5BoxB in different groups were calculated by the ratio of relative FL-5BoxB activities to their mRNA levels. In HEK 293 cells, tethered Ago2 did not significantly alter the FL-5BoxB reporter translation efficiency. However, tethered TNGW1 and GW182 reduced the translation efficiency FL-5BoxB reporter by 57.5% and 54.0%, respectively (^*P<0.01, t-test). Error bars represent standard deviations.