Each vial contains a mixed stable population of 500,000 Incucyte® Nuclight Green HT-1080 (Cat. No. 4486) and 500,000 Incucyte® Nuclight Red A549 (Cat. No. 4491). Parental HT-1080 and A549 cells were purchased from ATCC (Cat. No. CCL-121 and Cat. No. CCL-185, respectively). HT-1080 and A549 cells were made individually using the Incucyte® Nuclight Green Lentivirus (Cat. No. 4475; EF1α, puromycin) and Incucyte® Nuclight Red Lentivirus (Cat. No. 4476; EF1α, puromycin), respectively, at an MOI of 3 (TU/cell) in the presence of 8 μg/mL polybrene following the standard infection protocol, resulting in a ≥70% transduction efficiency. Individual cell lines were mixed 1:1 to a total final concentration of 1 million cells/mL in 90% FBS and 10% DMSO. All individual cell lines in our Incucyte® Nuclight catalog have been certified mycoplasma free by ATCC.