Establishing Bioluminescent and Fluorescent Cell Lines

HER2 and luciferase/GFP-expressing NIH3T3 cells (3T3/HER2-luc-GFP) were established with a transfection of RediFect Red-FLuc-GFP (PerkinElmer, Waltham, MA, USA). High GFP and luciferase expression was confirmed with 10 passages. Balb/3T3 cells stably expressing RFP were established with transfection by RFP (EF1a)-Neo lentiviral particles (AMSBIO, Cambridge, MA, USA). High RFP expression was confirmed in the absence of a selection agent with 10 passages. 3T3 cells stably expressing RFP (3T3-RFP) were used as negative controls [13 (link)]. Cells were grown in RPMI 1640 (Life Technologies, Gaithersburg, MD, USA) supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin (Life Technologies) in tissue culture flasks in a humidified incubator at 37°C at an atmosphere of 95% air and 5% carbon dioxide.