Figure 3.

The amount of Npi1 Ub ligase is reduced in npi1 cells. Crude extracts were prepared from cells grown on proline medium, resolved by electrophoresis, and blotted onto a nitrocellulose membrane. The blot was probed with polyclonal Nedd4 antibodies (Kumar et al. 1997). The strains used were 24346c (wild type; lane 1), 27038a (npi1; lane 2), and 27038a (npi1) transformed with plasmid YEpJYS-2 carrying the NPI1 gene (lane 3). The signal corresponding to Ub ligase Npi1/Rsp5 is marked with an asterisk.