FIG. 6.

Pap1 is ubiquitinated in vivo. (A) Extracts from W303 cells overexpressing an HA-Ub fusion were used for immunoprecipitation (IP) with anti-β-galactosidase (α β-gal) (lanes 1 and 3) or with the C-terminal Pap1 antibody (lanes 2 and 4), and the immunoblot was probed with the C-terminal Pap1 (lanes 1 and 2) or anti-HA antibodies (lanes 3 and 4). (B) Immunoprecipitated Pap1 from W303 cells overexpressing HA-tagged ubiquitin (lanes 1 and 3) or HA-tagged Smt3 (lanes 2 and 4) was probed with anti-Pap1 (lanes 1 and 2) or anti-HA (lanes 3 and 4) antibodies. The positions of molecular mass markers (in kilodaltons) are indicated to the left of the gels.