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. 2002 Nov;76(22):11612–11622. doi: 10.1128/JVI.76.22.11612-11622.2002

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Copyright © 2002, American Society for Microbiology

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FIG. 5. — The p32-binding domain is required for ORF73-dependent transactivation. Cos-7 cells were transfected with pSV40CAT (a), pRSVCAT (b), p73Δ4 (c), or pAWCAT1 (d) in the presence of pGFP, p73GFP, p73GFP+p32, p32, p73ΔNLS1, p73ΔNLS1+p32, p73ΔNLS2, or p73ΔNLS2+p32. The cells were harvested at 48 h posttransfection, and cell extracts were assayed for CAT activity. Results are shown in graphical form, where CAT activity is presented as fold induction over background levels. The variations between two replicated assays are indicated. (e) (i) Cos-7 cell extracts, untransfected (lane 1) or transfected with p73GFP (lane 2), p73ΔNLS1 (lane 3), or p73ΔNLS2 (lane 4), were resolved by SDS-PAGE, and GFP was detected by Western blotting using GFP antiserum. (ii) Cos-7 cell extracts transfected with p73GFP (lane 1), p73GFP+p32 (lane 2), p73ΔNLS1+p32 (lane 3), or p73ΔNLS2+p32 (lane 4) were resolved by SDS-PAGE, and p32 was detected by Western blotting using p32 antiserum.