FIG. 2.

Western analysis of in vitro packaging cell extracts. Extracts were prepared as described in Materials and Methods from 293 cells infected with adenovirus (Ad) alone or infected with adenovirus plus transfected with either dl63-87 or pIM45 plasmid DNA. Partial depletion of Rep proteins from the pIM45 + Ad extract (Rep-depleted lanes) was accomplished by incubating the extract with mouse anti-78/68 monoclonal antibody coupled to protein G-Sepharose beads. Ten microliters of each extract was electrophoresed on a polyacrylamide gel, transferred to a nitrocellulose membrane, and probed for Rep proteins (left side) by using the mouse anti-52/40 monoclonal antibody, which recognizes all four Rep proteins or for capsid proteins (right side) by using the guinea pig polyclonal anti-capsid antibody, which recognizes all three capsid proteins.