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. 2021 Sep 3;13(9):1760. doi: 10.3390/v13091760

Figure 4.

Figure 4

Effect of H5N1 NS1 and PA-X amino acid changes on IFN responses. (A,B) Human HEK293T cells were transiently co-transfected with a pCAGGS empty plasmid or with pCAGGS plasmids expressing the indicated HA epitope-tagged PA-X variants, together with a plasmid expressing Fluc under an ISRE dependent promoter, and a pCAGGS plasmid constitutively expressing Rluc. PA-XCIR protein encodes the amino acid changes T20A, A85T, T118I, I127V, G209E, V212A, L221R, and R250Q. PAOR-XCIR encodes the amino acid changes G209E, V212A, L221R, and R250Q. PACIR-XOR encodes the amino acid changes T20A, A85T, T118I, and I127V. At 24 h.p.t, cells were infected with SeV (Cantell strain, MOI 3) or left mock-infected (M), and 20 h later expression of Fluc (A) and Rluc (B) was analyzed by luminescence. Data show the mean and standard deviations from cells in triplicate. ns, non-significant; *, p < 0.05; **, p < 0.005; ***, p < 0.0005; ****, p < 0.0001 using a Student’s t-test. Experiments were repeated 3 times in triplicate, with similar results. R.u., relative units.