Cultivation of Mammalian Cell Lines

The HEK293T and HCT116 cells were obtained from ATCC (ATCC, Manassas, VA, USA). HCT116 cells with endogenous C-terminal NRF2 tagged with Nanoluc luciferase were generated in-house and have been previously described [23 (link)]. HUH-7 cells were procured from Xenotech (Kansas City, KS, USA). Both HEK293T and HUH-7 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) and Normocin™ (Invivogen, San Diego, CA, USA) as recommended by the manufacturer to prevent bacteria, mycoplasma, and fungal contamination. HCT116-NRF2-Nanoluc cells were maintained in McCoy’s medium enriched with 10% FBS and Normocin™. All cells were cultivated in a mammalian cell incubator at 37 °C and 5% CO₂.