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. 2012 Sep 19;7(9):e45730. doi: 10.1371/journal.pone.0045730

Figure 7. Characterization of NSs encoding alanine substitutions at the C-terminus.

Figure 7

(A) Schematics of NSs mutations. Acidic residues, Glu (E) at aa.263–255 and Asp (D) at aa.257–259, were substituted with alanine, and recombinant MP-12 encoding NSs mutations were recovered (NSs-E253-255A/D257–259, NSs-D257-259A and NSs-E253-255A). Underline indicates the amino acids substituted. (B) VeroE6 cells were mock-infected or infected with indicated viruses at an moi of 3. Cells were collected at 16 hpi, and the abundance of PKR (anti-PKR antibody), NSs and N (anti-RVFV antibody), and β-actin (anti-actin antibody) were analyzed by Western blot. (B) A549 cells were mock-infected or infected with indicated viruses at an moi of 3. Total RNA was extracted at 7 hpi, and IFN-β mRNA, ISG56 mRNA and RVFV anti-viral-sense S-RNA/N mRNA were detected by Northern blot with specific RNA probe. Representative data from at least 3 independent experiments are shown.