Qc6352

HepG2-NTCP, HepAD38, Huh1, PLC/PRF5, and Hep3B cells were maintained in DMEM (ThermoFisher) containing GlutaMAX plus 10% FBS, 50 U/ml Penicillin/Streptomycin, and non-essential amino acids. Plasticware was pre-treated with collagen (Sigma) and maintained at 37 °C with 5% CO₂/18% oxygen. Hypoxic treatments were performed in a hypoxic chamber (3%, or 1% oxygen) (Invivo 400, Baker-Ruskinn Technologies). JIB-04, QC6352, CPI-455 and GSK-J4 were purchased from MedChem Express.

IOX1,
IOX2, ML324, KDMOAM25, succinate,
fumarate, 2OG, dimethylpyridine-2,4-dicarboxylate, dimethyl 2-oxoglutarate,
dimethyl succinate, dimethyl fumarate, and DMOG were purchased from
Sigma-Aldrich, while IOX3, IOX4, Roxadustat, Vadadustat, Panobinostat,
disodium 2S/2R-hydroxyglutarate, and FG0041 were
obtained from Cayman Chemical Company. 2,4-PDCA (Alfa Aesar), Daprodustat
(MedChemExpress), QC6352 (MedChemExpress), Molidustat (Selleck Chem),
JIB-04 (Tocris Bioscience), GSK-J1 (Tocris Bioscience), GSK-J4 (Tocris
Bioscience), SD70 (Xcess Biosciences), CPI455 (Axon Med Chem), 2′-deoxy-5-methylcytidine
(TCI Chemicals), 2′-deoxy-5-(hydroxymethyl)cytidine (TCI Chemicals),
and (R/S)-Octyl-α-hydroxyglutarate
(Cambridge Bioscience) were purchased from other commercial sources.
All purchased inhibitors were >95% pure as determined by the manufacturer.
Key compound purity was verified using analytical HPLC using a UV
detector (210 or 254 nm) or CAD detector (Figures S18–S28). NOG was synthesized as described in the materials
and methods and has a purity >95% as determined by HPLC. All buffer
components were from Sigma-Aldrich unless otherwise stated.