Figure 6

PI3K is critical for WNV-activated nuclear translocation of IRF7. BMDMs were left untreated (mock) or infected with WNV in the presence of vehicle control (DMSO) or 5 μM of LY294002 (LY) for 24 h. (A) Immunoblotting analyses of IRF3, IRF7, and NF- κBp65 in the cytoplasmic (Gapdh as the housekeeping protein control) and nuclear fractions (Mcm2 as the housekeeping protein control). Blots shown are cropped from full-length. (B) Quantification of the nuclear immunoblot band density in (A) by Image J. The results are expressed as the ratio of IRF7/IRF3/p65 to Mcm2. (C) Immunofluorescence staining of IRF7 (red) and nuclei by DAPI (blue). Images were acquired with a Zeiss fluorescence microscope. The arrows indicate nuclear IRF7 staining. The data shown are representative of 3 independent reproducible experiments.