Luciferase reporter gene assay. NuLi-1 cells, MS-1 cells, A549 cells, and LK-2 cells were plated onto 24-well plates and incubated for 20 h at 37°C in 5% CO2. The control pGL3 vector DNA (0.2 μg) or similar amounts of Luciferase-NF-κB reporter vectors, was cotransfected into cells with the pRL-SV40 vector using the effectene transfection reagent. Cells were then treated with DMSO (0.016%, v/v), TNF (10 ng/ml), 17-DMAG (0.05 μM), or TNF (10 ng/ml) plus 17-DMAG (0.05 μM) for 24 h. Luciferase activity was measured in cell lysates. Each experiment was repeated thrice with similar results. *, it is considered as a significant difference, when P < 0.05 vs. corresponding control.
Qu et al. Diagnostic Pathology 2013 8:70 doi:10.1186/1746-1596-8-70