Dual nano glo dual luciferase assay kit

Cells were plated at 4.10⁵ cells/ml per well in a 96-well plate and co-transfected with nanoluciferase post-transcriptional reporter containing the ATR 3′UTR sequences and control non-ARE 3′UTRs that was fused with firefly luciferase. The cells were transfected using Lipofectamine 2000 (Invitrogen) according to the manufacturer’s instructions. Transfections were performed in several replicates. After 16 h, cells were lysed and assayed for nanoluciferase activity using the dual Nano-Glo dual-luciferase assay kit (Promega) according to the manufacturer’s instructions and measured on a luciferase luminometer. Data were presented as mean ± SEM of normalized Nano-luciferase intensity/firefly intensity.

Cells were plated at 2 × 10⁵ cells/mL per well in 96-well plate overnight and co-transfected with RPS30 promoter-linked nanoluciferase fused with the 3′UTR containing WT and mutant sequences along with firefly luciferase vector as a normalization control. The cells were transfected using Lipofectamine 2000 (Invitrogen) according to the manufacturer’s instructions. Transfections were performed in several replicates. After 16 h, cells were lysed and assayed for luciferase activity using the dual Nano-Glo dual-luciferase assay kit (Promega) according to the manufacturer’s instructions and measured on a luciferase luminometer. Data were presented as mean ± SEM of normalized Nano luciferase intensity/firefly intensity.