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. 2016 Jul 28;8(8):209. doi: 10.3390/v8080209

Figure 2.

Figure 2

NXT1 knockdown reduces viral replication kinetics. (A) Madin-Darby canine kidney (MDCK) or A549 cells were transfected with control or NXT1 siRNA for 24 or 48 h. The cells were subjected to WB analysis with anti-NXT1 and anti-β-actin antibodies (upper panel) or water-soluble tetrazolium (WST-1) assay (lower panel). Data shown are the results from two independent experiments and the mean values. (B) MDCK or A549 cells were transfected with control or NXT1 siRNA for 24 h and infected with influenza A/WSN/33 virus at a multiplicity of infection (MOI) = 0.0001 for 18 h and 48 h, respectively. The cells were then collected, lysed, and subjected to WB analysis with anti-WSN, anti-NXT1, anti-CRM1, and anti-β-actin antibodies. (C) MDCK or A549 cells were transfected with control or NXT1 siRNA for 24 h or 48 h, and infected with influenza A/WSN/33 virus at MOI = 0.0001. Supernatants were collected at 14, 18, and 22 h post-infection (hpi) from MDCK cells, or at 24, 48, and 72 hpi from A549 cells for virus titration by plaque assay. Viral replication kinetics were plotted for virus titer and time post-infection. Data shown are the results from two independent experiments (error bars) and the mean values.