Replication properties of the AIV and pH1N1 viruses in mammalian and avian cell types.

<p>A549, MDCK, and CEF cells were infected either with the H1N1/WSN (♦), H9N2 (▴), H5N2/F118 (▪) or H5N3 (○) viruses using an MOI = 4 and incubated at 37°C. (A) At hourly intervals post infection the cells were harvested and the vRNA levels quantified using qPCR as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033732#s3" target="_blank">methods</a>. Each value at a specific time point represents the mean of triplicate measurements (p<0.05). The data presented are a representative data set from one of two independent experiments. (B) At 9 hpi mock (M) infected cells or cells infected with either of the four viruses were radiolabelled for 1 hr in DMEM minus methionine (Invitrogen, USA) containing 100 µCi/ml [³⁵S] methionine (Perkin-Elmer, USA). Cells were extracted in boiling mix and analysed by SDS-PAGE. Protein bands corresponding to the neuraminidase protein (NA)/nucleoprotein (NP), the matrix (M) protein and nonstructural 1 (NS1) protein, and the uncleaved heamaggultinin (HA) are indicated. (C) Cells were infected with either the pH1N1/276 (♦), pH1N1/471 (▪), pH1N1/478 (▴), pH1N1/527(X) or H1N1/WSN (+) viruses, and at hourly intervals post infection the cells were harvested and the vRNA levels quantified using qPCR as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033732#s3" target="_blank">methods</a>. Each value at a specific time point represents the mean of triplicate measurements (p<0.05). The data presented are a representative data set from one of two independent experiments.