Fig 1: The effect of cap-analog, eIF4A(R362Q), and hippuristanol on translation of reporter mRNAs in vitro and in HeLa cells.(A–C) 0.1 pmol C+A+ reporter Firefly luciferase mRNAs with indicated 5′ UTRs were translated in Krebs extract in the presence of increasing concentrations of cap-analog (0.005, 0.02, 0.05, and 0.1 mM) or without it (A), or recombinant eIF4A(R362Q) (0.05, 0.1, 0.2 and 0.4 nmol/ml) or without it (B), or hippuristanol (1, 2, and 4 μM) or without it (C). Reaction mixtures were assayed for Firefly luciferase after 45 min incubation at 30 °C. The FLuc activity without addition of cap-analog (A) or eIF4A(R362Q) (B), or hippuristanol (C) was taken to be 100%. Values are the means of at least three independent experiments. Errors are 2 standard deviations. (D) Untreated or treated with hippuristanol during 1 h (0.1, 0.2, 0.4, and 0.8 μM) HeLa cells were transfected by reporter Firefly luciferase mRNAs with indicated 5′ UTRs and Renilla luciferase mRNA (as internal control), cultivated for 2 h, harvested and assayed for Firefly and Renilla luciferase (FLuc and RLuc, respectively). The FLuc/RLuc ratio for the control (untreated cells) was taken as 100%. Values are the means of at least three independent experiments. Errors are 2 standard deviations.