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. 2018 May 1;8:127. doi: 10.3389/fcimb.2018.00127

Figure 1.

Figure 1

Construction of the NS1 mutant IAVs in RIG-I-knockout 293T cells. The wild-type (WT) and RIG-I-knockout 293T cells were infected with Sendai virus (SeV) for 12 h and lysed for western blot analysis (A). The 12-plasmid reverse genetics system was used to rescue recombinant influenza viruses A/WSN/33 (WSN) expressing WT or mutant NS1 (R38A, K41A, or R38A/K41A) in 293T and RIG-I-knockout 293T cells. MDCK cells were then infected with the recombinant viruses rescued from the RIG-I-knockout 293T cells (MOI = 0.01) for 72 h and lysed for western blot analysis (B). NP and M1 were detected with their respective antibodies, and β-actin was probed as the loading control. Virus titers (C) in supernatants from the infected MDCK cells were tested at the indicated time points. Growth curves were performed in triplicate. Error bars represented the standard deviations from the mean values of the three independent assays.