Fig. 1.

RNA binding and inhibition of in vitro translation by human IFIT1. (A) EMSAs between human IFIT1 and capped-RNA visualized by SYBR Gold staining. Cap0-MHV, first 41 nt of MHV strain A59; Cap0-HCoV, first 42 nt of HCoV strain 229E; Cap0-GGG42, ACU to GGG modification of HCoV. The RNA secondary structure minimum free energy (kcal/mol) and 5′-overhang length (ovg) are indicated (see also Fig. S1B). (B) Schematic of bicistronic mRNA reporter. (C) Translation assay with IFIT1 titrated into Krebs extracts programmed with Cap0/m7Gppp reporter, and titration following a 10-min preincubation of the reporter with extracts. FF and Ren luciferase (luc.) activities at each concentration were normalized against buffer control, which was set to 1. Data represent the mean of two independent measurements performed in duplicate ± SD.