Steadylite plus luciferase substrate

Lipofectamine LTX (ThermoFisher Scientific, Waltham, MA, USA) was used according to the manufacturer’s recommendations to transfect HeLa cells with each FLuc-int-PTC construct. The cells were distributed 24 h later into white 96-well plates (Corning, Corning, NY, USA) and incubated for 20 h at 37°C and 5% CO2 with extract at 10 ng/μl or with G418 at 1 μg/μl. Luciferase activity was then measured by adding the Steadylite plus luciferase substrate (Perkin Elmer, Waltham, MA, USA) to each well and by measuring the visible light from each well for 10 sec with a Tristar luminometer (Berthold technologies, Versailles, France). The plate was read twice and the reported values are averages of the two reads.

HeLa cells were transfected with the FLuc-int-UGA construct using lipofectamine 3000 (Life technologies) according to the manufacturer’s recommendations. The cells were distributed 24 h later into white 96-well plates (Corning, Corning, NY, USA) and incubated for 20 h at 37 °C and 5% CO₂ with DMSO, DAP, or G418 at 1 mg/ml. Steadylite plus luciferase substrate (Perkin Elmer, Waltham, MA, USA) was then added to each well before measuring the visible light from each well for 10 s with a Tristar luminometer (Berthold technologies, Versailles, France). The plate was read twice and the reported values are averages of the two reads.