High-throughput Drug Screening in HepG2 Cells

HepG2 or HB214 (1500 cells/well) was seeded on a 384 well plate in 30 μl. Drugs were added after 24 h. Cell Titer Glo (CTG) viability assay was conducted after 72 h of drug treatment. Briefly, CellTiter-Glo 2.0 (#G9243 Promega) was added to each well in a 1:1 v/v ratio. Plates were then covered to keep from light and incubated at RT on an orbital shaker at 150 RPM for 30 mins. After the incubation, plate was read on a synergy H4 plate reader for luminescence. Dose effect curves for each drug were calculated using Prism software, version 9 (GraphPad). For drug combinations, responses were analyzed using SynergyFinder2.0^{50 (link)}. Drugs used include cisplatin (#479036-5 G, Sigma Aldrich), gemcitabine (#AC456890010, Fisher Scientific), vincristine (#AAJ60907MA, Fisher Scientific), triapine (#50-136-4826, Fisher Scientific), MK1775 (#M4102, LKT laboratories, Saint Paul, Minnesota), doxorubicin (#BP25161, Fisher Scientific), sorafenib (#NC0749948, Fisher Scientific), SN-38 (#S4908-50MG, Selleck Chemicals, Harris County, Texas), deferoxamine mesylate (#AC461770010, Acros Organics, Geel, Belgium), KU60019 (#S1570, Selleck Chemicals). All concentrations were seeded in triplicate and the experiment was repeated three times. Significance was determined using the Extra Sum of Squares f test.