FIG. 2.
Cdc48p is coprecipitated with 6myc-Hmg2p in yeast. The indicated yeast strains KFY100 (wild-type CDC48) and KFY194 (cdc48-10ts mutation), transformed with plasmid pRH244 encoding 6myc-Hmg2p (panels A and B, lanes 1 and 2 and lanes 4 and 5; panel C, lanes 1 to 4; panel D, lanes 3 to 5) or mock-transformed (mock) (panels A and B, lanes 3 and 6; panel C, lane 5; panel D, lanes 1 and 2), were grown for 3 h at the indicated temperatures. The 6myc-Hmg2p and associated proteins were precipitated with anti-myc antibodies (IP: anti-myc). Proteins were resolved by SDS-PAGE and were electroblotted onto nitrocellulose. For comparison, total cell extracts (1% of the proteins used for immunoprecipitation) (panel C, lanes 1 to 5) were resolved by SDS-PAGE and were electroblotted onto nitrocellulose. Blots in panels A, C, and D were probed with rabbit anti-Cdc48p followed by HRP-conjugated anti-rabbit IgG (IB: anti-Cdc48p). (B) The blot in panel A was reprobed with mouse anti-myc antibody followed by HRP-conjugated anti-mouse IgG (IB: anti-myc).