Fig. 2. Detection of Gcs1p and mutated forms in yeast by western blot analysis. The indicated amounts of proteins from (A) YPH 252 wild-type or gcs1 mutant cells and (B) purified His-tagged Gcs1p or (C) 
20 µg of proteins from gcs1 mutant cells overexpressing wild-type Gcs1p, HA-tagged zinc-finger-mutated (
Zn) Gcs1p, PH domain-deleted (PH) Gcs1p or GFP-fused PH domain (GFP-PH) were separated by SDS-PAGE. Proteins were stained with Coomassie blue (A) or transferred to PVDF, reacted with affinity-purified antibodies against Gcs1p and detected using the ECL system. Positions of protein standards (kDa) are at the left.
