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E1A binds directly to E4orf3. E. coli-expressed and -purified 6×His-E1A289R was mixed with either GST-E4orf3 or GST (also bacterially expressed and purified) and glutathione-agarose beads, incubated, washed, and resolved by SDS-PAGE. Pulldowns were blotted for E1A using M73 monoclonal antibody. Inputs for GST, GST-E4orf3, and 6×His-E1A289R (1 µg of each protein) were resolved by SDS-PAGE and stained with Coomassie. L, molecular weight ladder.
