FIG. 5.

(A) Supplementation of immunodepleted extracts. Replication assays of AAV were performed as described in Materials and Methods in a mock-depleted extract from Ad-infected cells (lane 1) and an MAb 37-3-depleted extract from Ad-infected cells (lanes 2 to 4). After depletion, extracts were unsupplemented (lanes 1 and 2) or supplemented with 1.5 μg of Ad-DBP (lane 3) or 1.0 μg of RPA (lane 4). (B) Supplementation of a mock-depleted extract. Replication of AAV was performed as described in Materials and Methods in a mock-depleted extract from Ad-infected cells. Lane 1, extract supplemented with RPA after mock depletion; lane 2, extract not supplemented. (C) Western blot of depleted and mock-depleted extracts from Ad-infected cells with an antibody to the 34-kDa subunit of RPA. Lane 1, depleted extract; lane 2, mock-depleted extract. Size markers, shown on the right, are in kilodaltons. Additional bands in lane 1 are residual antibody from the immunodepletion, detected by the secondary antibody in the detection system (goat anti-mouse IgG).