Human NSCLC lines A549, H460, H1944, H1792, H23, H1299, H1975 and H1650 were purchased from LGC Ltd. H460 FLuc were purchased from PerkinElmer. H1299 FLuc were purchased from AMS biotec. A549 NRF2 knockout (KO), A549 KO-restored (KO-R), H1299 T80K and H1299 PLX317 were previously described 24 , 25 (link). To pharmacologically activate NRF2 expression, cells were treated with 100 nM of the KEAP1 inhibitor, KI696, for 24 h. All cells were cultured in RPMI media (Sigma-Aldrich), supplemented with 10% foetal bovine serum (ThermoFisher Scientific) and 100 U.mL−1 penicillin and 100 μg.mL−1 streptomycin (Sigma-Aldrich), maintained at 37 °C and 5% CO2. Cell lines were tested monthly for mycoplasma infection (Eurofins).
Cells were seeded for 24 h prior to experimental endpoint in 6-well plates in 2 mL of media. All cells were seeded at 1.5 × 105/mL, except for H1944 and H1792, which were seeded at 2.75 × 105/mL; H1650, which was seeded at 1.75 × 105/mL; and H23 and H1975, which were seeded at 2.5 × 105/mL. For KI696 treatments, H1650, H23, H1975 and H1299 were seeded at 1.0 × 105/mL, 1.0 × 105/mL, 1.25 × 105/mL and 0.75 × 105/mL, respectively 2 mL of media 24 h prior to treatment.
Cells were seeded for 24 h prior to experimental endpoint in 6-well plates in 2 mL of media. All cells were seeded at 1.5 × 105/mL, except for H1944 and H1792, which were seeded at 2.75 × 105/mL; H1650, which was seeded at 1.75 × 105/mL; and H23 and H1975, which were seeded at 2.5 × 105/mL. For KI696 treatments, H1650, H23, H1975 and H1299 were seeded at 1.0 × 105/mL, 1.0 × 105/mL, 1.25 × 105/mL and 0.75 × 105/mL, respectively 2 mL of media 24 h prior to treatment.