FIG. 3.

Western blot analysis of HA-tagged Lac1p and Lag1p. Whole-cell protein extracts were prepared from the indicated strains, and equal amounts of each extract were electrophoresed by SDS-PAGE. Each strain contained a low-copy-number plasmid expressing a 3× HA-tagged form of either Lac1p (top) or Lag1p (bottom). Where indicated, a low-copy-number plasmid carrying hyperactive PDR3 (PDR3-11) or the empty vector (pRS315) was also present. After electrophoresis, the proteins were transferred to nitrocellulose and the membrane was probed with an anti-HA antibody. A monoclonal antibody directed against the vacuolar membrane protein Vph1p was used to ensure equal loading and transfer.