Figure 5.

Comparison of translation of unmodified and m⁶A-modified reporter mRNA in the presence of eIF4A (R362Q) (A) or hippuristanol (B). C + A + Firefly luciferase reporter mRNA carrying the beta-globin 5′ UTR (0.15 pmol) was translated in HEK293T cell extract with or without increasing concentrations of recombinant eIF4A (R362Q) (0.43, 0.87, 1.3, and 1.74 µmol) or increasing concentrations of hippuristanol (1, 2, and 4 µmol). The reaction mixture was incubated for 60 min at 30 °C before the determination of Firefly luciferase activity, which was taken to be 100% with no eIF4A (R362Q) or hippuristanol added. The results of three independent experiments are presented. Bars are 2 standard deviations. A two-tailed Student’s t-test was used to estimate the statistical significance; ** p < 0.01; *** p < 0.001.