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Figure 5 | Veterinary Research

Figure 5

From: Identification of two residues within the NS1 of H7N9 influenza A virus that critically affect the protein stability and function

Figure 5

I178V mutation decreases stability of H7N9 NS1 protein. A 293T cells were transfected with plasmids expressing H7N9 NS1-WT (WT) or NS1 mutants (R55E, H63Q, E70K, P87S, S114P, A143T, I178V, S212P) for 24 h, followed by Western blotting to detect the NS1 protein levels. The results are representative of three independent experiments. B 293T cells were transfected with plasmids expressing H7N9 NS1-WT (WT) or NS1-I178V (I178V) as described in (A). Then the mRNA levels of NS1 were detected by RT-PCR. C Half-lives of NS1-WT (WT) and NS1-I178V (I178V) were examined. 293T cells were transfected with plasmids expressing H7N9 NS1-WT (WT) or NS1-I178V (I178V) as described in (A), then the cells were treated with CHX (100 μg/mL). At the indicated times after treatment, cells were harvested and cell extracts were prepared for Western blotting to analyze NS1 protein levels. D NS1 levels in (C) were quantitated by densitometry, and normalized to β-actin levels. Plotted are the average levels from three independent experiments. *P < 0.05, **P < 0.01.

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