Two × 106 HFFs were plated in 6 well plates, and the following day the cells were infected with OC43 (MOI = 1) for 1 h. Cells were then washed with PBS once, and OZ418 (30 μM), OZ277 (15 μM), AS (30 μM), emetine (200 nM), or DMSO was added to infected cells. All drugs were diluted in DMEM containing 4% FBS along with noninfected and infected nontreated (DMSO only) controls. At 72 h postinfection, the cells were washed with PBS and lysed in cell lysis buffer (Promega) containing protease (Roche) and phosphatase (Thermo Scientific) inhibitors. Lysate from each condition (50 μg) was separated on 10% SDS-PAGE for detection of the OC43 antigen, and 25 μg of lysate from each condition was analyzed for β-actin (Sigma, 1:10,000) as the loading control. Anti-coronavirus monoclonal antibody, OC-43 strain, clone 541-8F (Merck Millipore, MAB9012) (1:250) was used to detect the OC43 antigen (not defined).

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