Stable Transfection of NSCLC Cell Lines

Non-small cell lung cancer cell line NCI-H1299 (ATCC, Manassas, VA, USA) was stably transfected with BB₁R, BB₂R, and BRS-3. The transfected cells were grown in RPMI-1640 containing 10% fetal bovine serum (FBS) with 0.3 mg/ml geneticin (Invitrogen, Grand Island, NY, USA). The transfected cells, which contained approximately 100,000 receptors/cell, were weekly split using trypsin/EDTA (13 (link)). In addition, lung cancer cell lines NCI-H727, H1299, and H1975 were purchased from ATCC and cultured in RPMI-1640, which contained 10% FBS. The cell types were derived from different human biopsy specimens. These studies were approved by the NIDDK biospecimens and biosafety committees.