All drugs tested in this study, nafamostat (Medchemexpress), remdesivir (Selleckchem), toremifene (Selleckchem), nelfinavir (Selleckchem), fenofibrate (Selleckchem), and clofzamine (Selleckchem) had a purity >95% and were dissolved in dimethyl sulfoxide (DMSO) to a stock concentration of 10 mM. The drug stocks were diluted in the DM culture medium and flowed through the apical epithelial channel (toremifene 10 μM, nelfinavir 10 μM, fenofibrate 25 μM, clofazamine 1 µM) or EM in the basal microvascular channel (nafamostat 10 μM, remdesivir 1 and 10 µM) based on whether they are normally administered orally or intravenously in humans, respectively. Treatment was started 1 day prior to viral infection and continued for 48–72 h as indicated in figure legends.
For studies on the effects of drugs on human umbilical vein endothelial cell (HUVEC; Lonza, C2519A) viability, the cells were expanded in EGM-2 medium with 2% FBS (Lonza) and plated at 10,000 cells/well in a 96 well plate. 72 h after plating, wells were treated with fresh medium alone, with medium with DMSO or remdesivir. 48 h after dosing, CellTiter-Glo (Promega #G7571) assay was used to measure cell viability.
For studies on the effects of drugs on human umbilical vein endothelial cell (HUVEC; Lonza, C2519A) viability, the cells were expanded in EGM-2 medium with 2% FBS (Lonza) and plated at 10,000 cells/well in a 96 well plate. 72 h after plating, wells were treated with fresh medium alone, with medium with DMSO or remdesivir. 48 h after dosing, CellTiter-Glo (Promega #G7571) assay was used to measure cell viability.