The four activators and the Swi/Snf complex occupy the STA1 promoter in the absence of glucose. (A) FLO8 and TEC1 expression is increased, but amounts of Nrg1 and Sfl1 are reduced under derepressing conditions. HA-tagged strains (wild type) were grown in synthetic medium containing 2% glucose (repressed condition [R]) or 2% glycerol-ethanol (derepressed condition [D]) to mid-log phase. Protein extracts (100 μg) prepared from each integrated HA-tagged strain were separated by SDS-PAGE and Western blot analysis with anti-HA (α-HA) antibody. The same membranes were probed with an anti-actin monoclonal antibody. (B) The four activators occupy the STA1 promoter under derepressing conditions. HA-tagged strains (wild type) were grown in synthetic medium containing 2% glucose (R) or 2% glycerol-ethanol (D) to mid-log phase, and anti-HA ChIP assays were performed as described above with 1 mg (for Flo8 and Mss11) and 500 μg (for Ste12, Tec1, Nrg1, and Sfl1) of total extracts to determine the binding of transcription factors to the STA1 promoter.