Skip to main content
. 2010 Jul 21;84(19):10051–10062. doi: 10.1128/JVI.00592-10

FIG. 5.

FIG. 5.

Lack of H5N1 PB1-F2 reduces polymerase activity in transfected cells. 293T cells were cotransfected with Pol I-Pol II system expression plasmids with PR8 PB2, PR8 PA, PR8 NP, and PR8 PB1 with or without PR8 PB1-F2 (A) or H5N1 PB1 with or without H5N1 PB1-F2 (B), together with a plasmid that enabled the expression of a pseudoviral reporter RNA (plasmid pPolI-CAT-RT). Vec ctrl, vector control, in which 293T cells were cotransfected with PR8 PB2, PR8 PA, PR8 NP, and pPolI-CAT-RT plasmids without PR8 PB1. At 48 h posttransfection, total cell lysate was extracted with lysis buffer and was assayed using a CAT ELISA. The optical density at 405 nm for PR8 PB1delF2 (A, top panel) or H5N1 PB1delF2 (B, top panel) was normalized according to that for PR8 PB1 (A, top panel) or H5N1 PB1 (B, top panel), which was considered 100%. Mean values and standard errors from three independent experiments are shown. **, P < 0.001, Student's two-tailed unpaired t test. (A, lower panel) Expression of PR8 PB1 and PR8 NP in 293T transfected cells, monitored in Western blots with polyclonal anti-PB1 and monoclonal anti-NP antibody; (B, lower panel) expression of H5N1 PB1 and PR8 NP in 293T transfected cells, monitored in Western blots with polyclonal anti-PB1 and monoclonal anti-NP antibodies.