Figure 3.

Deletion of SLA1 affects Rho1p localization. (A and B) Immunoblots of whole-cell lysates (10 μg/lane) probed with affinity-purified anti-Rho1p antibodies. (A) Lysate of wild-type cells (DDY664) probed with untreated antibodies (left lane) or with the same antibodies after preincubation with the Rho1p peptide antigen (right lane). (B) Lysates of DDY1097 (carrying a GAL–RHO1 plasmid; right lane) and DDY1098 (carrying a control plasmid) were prepared after growth on galactose for 8 h and probed with the anti-Rho1p antibodies (bottom) or (as a loading control) with anti-β-tubulin antibodies (top). (C and D) Immunolocalization of Rho1p in log-phase cells of wild-type (C) and sla1 null cells (D) Bar, 5 μm. (E) The staining patterns of unbudded cells were counted. The data shown are the averages of three experiments in which at least 200 unbudded cells were scored. The single patch of staining in wild-type unbudded cells is presumably the site from which the new bud will emerge.