FIG 8.

MCM-BP binds USP7 on chromatin and can mediate an interaction between the USP7 and MCM proteins. (A) Immunoprecipitations were performed with soluble and chromatin-associated lysates from HeLa cells using antibody against MCM-BP or negative-control IgG. Immunoprecipitants were analyzed by SDS-PAGE and Western blotting against the indicated proteins. (B and C) FLAG-tagged MCM7 (B) or FLAG-tagged MCM5 (C) was expressed individually or coexpressed with MCM-BP (BP) or MCM-BP S158A (S158A-BP) in insect cells. In each case, these proteins were expressed with and without USP7, as indicated. Insect cell lysates were then incubated with anti-FLAG resin to recover FLAG-MCM7 or FLAG-MCM5 with associated proteins. Bound proteins were eluted with protein sample buffer and analyzed by SDS-PAGE and Coomassie staining (top panel) or Western blotting for USP7 (bottom panel).