1. Arginine-to-lysine mutations conferring TRIM22 restriction and lysine-to-arginine mutations causing loss of TRIM22 restriction.
Arginine-to-lysine mutations conferring TRIM22 restriction and lysine-to-arginine mutations causing loss of TRIM22 restriction. TRIM22-mediated restriction activity was tested in the polymerase activity assay in the presence of 40 ng of TRIM22-expressing plasmid. (A) The bars show the activity of WT pH1N1 NP and R-to-K mutants relative to the same samples tested in the absence of TRIM22-expressing plasmid, which was set to 100%. Means ± SD from three independent experiments performed in triplicate are reported. The results of one-way ANOVA with the Bonferroni’s multiple-comparison test of WT versus each mutant are shown (*, P < 0.05; ****, P < ; ns, not significant). The expression levels of pH1N1 NP and TRIM22 were determined by Western blotting of WCE obtained by pooling the triplicate wells of each experiment. One representative Western blot of three is shown. (B) The bars show the activity of sH1N1 NP WT and K-to-R mutants relative to the same samples tested in the absence of TRIM22-expressing plasmid, which was set to 100%. Means ± SD from three independent experiments performed in triplicate are reported. The results of one-way ANOVA with the Bonferroni’s multiple-comparison test of WT versus each mutant are shown (****, P < ). Western blot analysis was performed to determine the expression levels of sH1N1 NP and TRIM22. (C) Quantification of the band intensity was performed by using ImageJ. Means ± SEM of the ratio between the intensity of NP bands and β-actin are reported for 3 independent experiments. The results of one-way ANOVA with the Bonferroni’s multiple-comparison test are shown (*, P < 0.05).
Isabel Pagani et al. mSphere 2018; doi: /mSphere