FIG 5.

The identified mutations affect nsp1-induced inhibition of cellular translation. (A) BHK-21 cells were transfected with plasmids encoding GFP-fusions with either wt nsp1 or its variants with the indicated mutations. At 18 h post transfection, cells were incubated for 15 min in the media supplemented with 10 μg/mL of Pur. Cell lysates were processed, as described in Materials and Methods. The top panel presents a Western blot stained with anti-Pur and corresponding secondary antibodies. The images were acquired on a LI-COR imager. (B) Quantitation of the Pur signal of the blots, one of which is presented in panel A. The means and SDs are presented. The statistical significance of differences was determined by one-way ANOVA with a post hoc Dunnett’s multiple-comparison test (n = 4).