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. 2007 Sep 12;81(22):12427–12438. doi: 10.1128/JVI.01105-07

FIG. 5.

FIG. 5.

Overexpression of HA-4E-BP1 proteins does not affect influenza virus infection in HEK293T cells. (A) HEK293T cells were untransfected (M) or transfected with plasmids expressing HA-tagged wild-type 4E-BP1 (WT) or nonphosphorylatable 4E-BP1 (4A) protein. At 36 h posttransfection, total cell extracts (T.ext.) were used to study eIF4GI, eIF4E, and HA-4E-BP1 (HA) retention either to cap resins or to Sepharose-4B control resins (ctrl-resin) by Western blotting. Quantitation of eIF4GI protein retained on the cap resins is shown on the right. (B) HEK293T cells were transfected with plasmid pcDNA3-HA-4E-BP1 wt or pcDNA3-HA-4E-BP1 4A, and 36 h posttransfection, the cells were mock or influenza virus infected with either the VIC (top) or delNS1 (bottom) strain. At the indicated hpi, cells were fixed and used for immunofluorescence using antibodies against HA to monitor plasmid transfection and NP protein to monitor influenza virus infection. Asterisks indicate transfected and infected cells. DAPI, 4′,6′-diamidino-2-phenylindole.