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. 2014 Aug;88(16):9038–9048. doi: 10.1128/JVI.00830-14

FIG 1.

FIG 1

Generation of a recombinant PR8 virus with epitope-tagged NS1- and NS2 (PR8-NS-FS). (A) IAV NS gene arrangement and structure of recombinant PR8-NS-FS virus through the duplication of 90 bp of the 5′ gene segment (hatched), followed by the 2A peptide of the porcine teschovirus-1 (P2A) and a tandem triple-FLAG-StrepOne epitope tag (FS). Different expected mRNA and protein products for PR8-wt (upper part) and PR8-NS-FS (lower part) are shown. Triangle, 2A-encoded “cleavage” site. (B) Replication of PR8-wt and PR8 NS-FS on MDCK, A549, and HEK293T cells was determined by infection at an MOI of 0.001 (MDCK and A549 cells) or at an MOI of 0.01 (HEK293T cells) and determination of the virus titer from the supernatants at the indicated time points. (C) Immunoblot analysis of MDCK, A549, and HEK293T cells, infected with PR8-wt (wt) and PR8-NS-FS (FS) at an MOI of 1 for 24 h. Blots were analyzed with antibodies against proteins indicated (left), and the protein identity is indicated (right).