2.1. Cell culture and transfection

HepG2 cells (ATCC HB‐8065), which are routinely used as a robust in vitro model for the liver in metabolic studies, 27 were cultured using DMEM/F‐12 growth medium with 10% fetal bovine serum, 1% (10 000 U/mL) penicillin/streptomycin, and 1% (110 mg/L) l‐glutamine at 37°C with 5% CO₂. Cells are passaged at 80% to 100% confluence using 0.25% Trypsin (wt/vol)‐0.53 nM ethylenediaminetetraacetic acid. For transfection, nearly confluent cells were passaged 24 hours prior, then transfected following the Lipofectamine 3000 (ThermoFisher #L3000001) protocol for 24 well plates using 500 ng of each plasmid and 1.5 μL Lipofectamine 3000 reagent per well. Transfection mixture was left on cells for 48 hours before imaging, and for 72 hours before protein harvest for western blot analysis.