Journal: PLoS ONE

Article Title: Effects of the C-Terminal Truncation in NS1 Protein of the 2009 Pandemic H1N1 Influenza Virus on Host Gene Expression

doi: 10.1371/journal.pone.0026175

Figure Lengend Snippet: In vitro characterization of the recombinant viruses encoding different-length NS1 protein. (A) A549 or PK-15 cells were infected at an MOI of 0.001 TCID 50 per cell. After washed with PBS, fresh medium containing 0.1 µg/ml TPCK-trypsin was added. Supernatants were sampled at 12, 24, 36 and 48 h p.i. and virus titers were determined as log 10 TCID 50 /ml in MDCK cells. The statistical analysis was performed using Student's t test. (B) Virus plaque assay was performed by inoculating confluent MDCK cells in 6-well plates with appropriate dilutions of viruses. After washed with PBS, cells were overlaid with DMEM (without phenol red)-0.8% agarose mixture containing 1% bovine serum albumin and 1 µg/ml TPCK-trypsin. After incubated at 37°C for 4 days, a second agar overlay containing 1∶10,000 neutral red was added.

Article Snippet: Virus plaque assay of the two recombinant viruses rPR8-Mex NS1/219 and rPR8-Mex NS1/230 was performed by inoculating confluent MDCK cells in 6-well plates with appropriate dilutions of viruses in DMEM for 1 h. Cells were washed twice with PBS and then overlaid with DMEM (without phenol red, HyClone)-0.8% agarose mixture containing 1% bovine serum albumin and 1 µg/ml TPCK-trypsin.

Techniques: In Vitro, Recombinant, Infection, Plaque Assay, Incubation