Fig 1. Polymerase activity of zoonotic avian influenza viruses in human cells.
Viral polymerase activity was measured by minigenome assays in human 293T cells. (A) Polymerase activity of zoonotic avian influenza viruses at 33°C. The data are expressed relative to the results for G1/1997 (H9N2). (B) Polymerase activity at 33°C of reassorted polymerase complexes composed of one subunit from EG/2013 (H9N2) and the other subunits from ancestral G1/1997 (H9N2). The data are expressed relative to the results for EG/2013-wt. (C) Western blot analysis of reassortant polymerase complexes shown in B without (Input) and with (IP: PB2-flag) immunoprecipitation with PB2-flag. Representative images are shown. (D) Polymerase activity of PB2-K627E revertants from EG/2007 (H5N1) and Sh/2013 (H7N9), and of the PB2-V627E revertant from EG/2013 (H9N2) at 37°C and 33°C. (E) Polymerase activity of the PB2-V627K mutant from EG/2013 (H9N2) at 37°C and 33°C. The values of the fold reduction relative to wild-type (D) and to the PB2-V627K mutant (E) are shown. (F) Quantification of PB2 expression levels in 293T cells. The cells were transfected with plasmids expressing the indicated non-tagged PB2, and cell lysates were analyzed by western blotting using anti-PB2 antibody. Representative images are shown. Each data point is the mean ± SD of three independent experiments. Asterisks indicate a P value < 0.01. NS indicates no statistically significant difference.
