Figure 5.
Slx5, but not Slx8, interacts physically with α2. Yeast ubc4Δ ubc6Δ matα2Δ cells (MHY3765) cells were cotransformed with pRS425-GAL1-α2 and a pYES2.1 (2 μm, GAL1) plasmid expressing V5-His6-tagged Slx5, Slx8, or Cdc48 (negative control). After induction with galactose, cells were lysed under native conditions, and V5-tagged proteins were immunoprecipitated with anti-V5-agarose. Slx5, Slx8, and Cdc48 proteins were detected by anti-V5 (Invitrogen), and coprecipitated α2 by anti-α2 immunoblotting. For lane 3, half of each of the number of cells used for lanes 4 and 5 were mixed and lysed together, so the amount of input Slx8 and Slx5 was half of that used for each of the proteins in lanes 4 and 5. (Lane 1) A control immunoprecipitation with cells that overexpressed α2 but lacked any V5-tagged protein.