Murine Lewis lung carcinoma (LLC) cells were purchased from ATCC® (American Type Culture Collection, Manassas, VA, USA) and cultured in either T-75 or T-175 flasks. Cells were passaged for subculturing by first aspirating the culture medium with a pipette, rinsing with 1× phosphate buffered saline (PBS, Thermo Fisher Scientific, Waltham, MA, USA, SH30256FS), aspirating off the PBS, then rinsing with 0.25% trypsin-0.53 mM EDTA solution (Thermo Fisher Scientific, Waltham, MA, USA, 25-200-056), and then they were neutralized with complete growth media consisting of Dulbecco’s modified Eagle’s medium (DMEM, ATCC®, Manassas, VA, USA) with 10% fetal bovine serum (FBS, USDA approved, ATCC®, Manassas, VA, USA), and 1% Penicillin-Streptomycin (10,000 U/mL, Thermo Fisher Scientific, Waltham, MA, USA). Cells were modified to be luciferase-expressing (LLC-Luc), as previously described [54 (link)] through use of plasmid pLenti PGK V5-LUC Neo [63 (link)] (Addgene, Cambridge, MA, USA) which was packaged in lentiviral particles and performed at the Baylor College of Medicine (BCM) vector core facility. For the luciferase-expressing cells, 1% Geneticin (Thermo Fisher Scientific, Waltham, MA, USA) was added to the media to maintain culture. Cells were maintained in a HERAcell 150i CO2 incubator (Thermo Fisher Scientific, Waltham, MA, USA) set to 37 °C and 5% humidity.
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