The FDA-Approved Oral Drug Nitazoxanide Amplifies Host Antiviral Responses and Inhibits Ebola Virus

Jasenosky, Luke D.; Cadena, Cristhian; Mire, Chad E.; Borisevich, Viktoriya; Haridas, Viraga; Ranjbar, Shahin; Nambu, Aya; Bavari, Sina; Soloveva, Veronica; Sadukhan, Supriya; Cassell, Gail H.; Geisbert, Thomas W.; Hur, Sun; Goldfeld, Anne E.

supplemented with 10% heat-inactivated fetal calf serum (FBS). The next day, cells were transfected with 10 ng of pFLAG-CMV4 plasmid encoding RIG-I, MDA5, or an empty vector control (Ahmad et al., 2018; Wu et al., 2014) , together with 100 ng of IFNβ promoter-driven firefly luciferase reporter plasmid and 10 ng of CMV promoter-driven Renilla luciferase reporter plasmid (Lipofectamine2000, Life Technologies). The medium was changed 4 hours post-transfection with DMEM containing 1% FBS and the indicated concentration of NTZ. All experiments involving NTZ were performed in this manner in order to reduce the impact of serum proteins on NTZ's function in tissue culture, as NTZ is highly protein-bound (Stockis et al., 1996) . Four hours after addition of NTZ (Sigma),