FIGURE 5.

Interaction of p97 with a heavy chain retrotranslocation intermediate in US2 cells. MG132-treated US2 cells were radiolabeled. Cells were lysed in buffers containing either 1% DeoxyBigCHAP (DBC) or 0.5% Nonidet P-40. Cell extracts were subjected to immunoprecipitation with either control IgG or αp97. A portion of the precipitated material was directly analyzed by SDS-PAGE and autoradiography (bottom panel). The remaining material was subjected to a second round of immunoprecipitation with αHC. Where indicated, a fraction of cell extract was subjected to immunoprecipitation with αHC antibody. Shown is a representative gel from two independent experiments.