H1299 human lung carcinoma cells (ATCC CRL-5803) were cultured in high glucose DMEM (Sigma- Aldrich, St. Louis, MO) with 10% FBS, 2 mM glutamine and 1× Penicillin and Streptomycin on plastic plates (BD biosciences, San Jose, CA) at 37 °C. Cells were seeded at 7 × 105 per well in 6-well plates (BD biosciences, San Jose, CA). 24 h after seeding, cells were transfected in complete media with 2 µg of test DNA, along with 135 ng of β-galactosidase plasmid and 1.1 µg of pUC19 DNA using 3 µl of Fugene HD per 1 µg of DNA. Cells were harvested 30 h after addition of DNA, and processed and assayed as described above. 15 µl and 2.5 µl of the extract were used for Luciferase and β-galactosidase assays, respectively; activity was measured on a Monolight 2010 Luminometer (Analytical Luminescence Laboratory, Sparks, MD).
