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.

To generate A549 cells with stable knock down of specific genes, cells were transduced with the control or gene-specific lentiviruses, and positively transduced cells were selected with 2 µg/ml puromycin for 3 weeks as described previously (Chow et al., 2014) . Efficacy of the knockdowns was then evaluated by analyzing gene expression following mock stimulation or stimulation with 100 ng/ml human IFN-β for 4 hours for the CRISPR-control, CRISPR-RIG-I, and CRISPR-PKR cells, or with 40 µM NTZ for 8 hours for the CRISPR-control and CRISPR-GADD34 cells. RNA was isolated and reverse transcribed using anchored oligoDT priming and cDNA was measured using standard qPCR methodology and normalized to the housekeeping gene cyclophilin B