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Fig. 3 | Virology Journal

Fig. 3

From: Influenza A virus NS1 optimises virus infectivity by enhancing genome packaging in a dsRNA-binding dependent manner

Fig. 3

Effects of NS1 on viral replication in the VLP system. a 293 T RIG-I KO cells were co-transfected with eleven expression plasmids as described and a minigenome expression construct. Minigenome expression is regulated by RNA pol I as illustrated. b Twenty-four hours after transfection, cells and VLP released in the culture medium were harvested. VLP was further purified (Materials and Methods). Proteins in donor cells and purified VLP were detected by immunoblotting. c Luciferase activity in the donor cells was measured. Firefly luciferase activity was normalised by Renilla luciferase activity. Luciferase activity without PB1 expression was set to 1. The Student’s t test was used for statistical analysis (n.s. not significant). The data shown are the mean ± standard deviation from at least three independent experiments (n = 3). d Total RNA was extracted from the donor cells and minigenome was quantified by strand specific RT-qPCR. Levels of firefly luciferase vRNA obtained from cells without PB1 expression were set to 1. The Student’s t test was used for statistical analysis (n.s. not significant). The data shown are the mean ± standard deviation from at least three independent experiments (n = 3). e vRNA in the purified VLP was quantified by strand specific RT-qPCR. Levels of Firefly luciferase vRNA obtained from E.C. transfected cells were set to 1. The Student’s t test was used for statistical analysis (*P < 0.05). The data shown are the mean ± standard deviation from at least three independent experiments (n = 3)

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