FIG. 2.

Electrophoretic mobility supershift assays and immunoprecipitation of WCEs with anti-FKBP52 antibody. The AAV D-sequence probe (lane 1) was incubated with WCEs prepared from HeLa cells to yield a slower-migrating complex (lane 2; solid arrow) and with those from 293 cells to form a faster-migrating complex (lane 3; solid arrowhead). These complexes were supershifted by incubation with antiFKBP52 antibody with HeLa (lane 4; open arrow) and 293 (lane 5; open arrowhead) WCEs, respectively, but not with anti-β₁ integrin antibody (lanes 6 and 7). No complex formation occurred between the AAV D-sequence probe and either anti-FKBP52 antibody (lane 8) or anti-β₁ integrin antibody (lane 9). When WCEs from HeLa and 293 cells were immunoprecipitated with anti-FKBP52 antibody and supernatants and pellets, following resuspension, were used in EMSA, prior immunoprecipitation with anti-FKBP52 antibody eliminated ssD-BP from WCEs from both HeLa and 293 cells (lanes 10 and 11), and it could be recovered from the pellets (lanes 12 and 13).