Primary human hepatocytes

Human hepatoma cells (HepG2) were maintained in DMEM supplemented with 10% fetal bovine serum (Hyclone). Primary murine hepatocytes were isolated and cultured as previously described.^[52 (link)
^] Primary human hepatocytes were purchased from Sigma. Human DDIT4 promoter–luciferase constructs^[53 (link)
^] and BRG1 expression constructs^[54 (link)
^] have been previously described. Small interfering RNAs were purchased from Dharmacon. Transient transfections were performed with Lipofectamine 2000. Luciferase activities were assayed 24–48 h after transfection using a luciferase reporter assay system (Promega).

The human medullary thyroid carcinoma cell line (TT) was procured from the American Type Culture Collection (Manassas, VA, USA) and cultivated in DMEM 1640 medium (Cellgro, Manassas, VA, USA) enriched with 15% heat-inactivated fetal bovine serum (FBS). The isogenic cell line HEK293-RET, containing the luciferase reporter gene regulated by the wild-type RET promoter sequence, was produced as detailed in our prior research [23 (link),28 (link)]. This cell line was cultured in DMEM 1640 media supplemented with 9% FBS. Dr. Rebecca Schweppe from the University of Colorado, Denver, supplied the TPC1 cells and the MZ-CRC-1 cells, which are derived from MTC. The TPC1 cells were cultured in RPMI-1640 medium with an added 9% of fetal bovine serum (FBS), while the MZ-CRC-1 cells were grown in DMEM/F-12 medium with a 15% supplementation of FBS. Primary Human Hepatocytes was purchased from Sigma Aldrich with its Human Hepatocyte Thawing Medium (MED-HTTM), Human Hepatocyte Culture Medium (MED-HHCM), Human Hepatocyte Plating Medium (MED-HHPMSP), and Supplement Pack (MED-HHCMSP). N-Thy-Ori 3-1 and K1 cell lines were grown in RPMI-1640 and Ham’s F12 medium (Thermo Fisher Scientific, Waltham, MA, USA) enriched with 9% FBS, respectively. All the cell lines were kept in a humidified environment containing 5% CO₂ at 37 °C.