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. 2004 Nov;24(21):9542–9556. doi: 10.1128/MCB.24.21.9542-9556.2004

FIG. 6.

FIG. 6.

Effects of Ste12-Tec1 on binding of Flo8-Mss11 to UAS1-2 in the artificial plasmid or endogenous STA1 promoter. (A) Effect of Ste12 and Tec1 on lacZ expression from the STA1p-lacZ plasmid. The STA1p-lacZ plasmid was transformed into the wild type (WT) and each mutant, and three independent transformants of each were tested for β-galactosidase (β-gal) activity under the derepressed condition (2% glycerol-ethanol). (B) Binding of Flo8 to UAS1-2 in the STA1p-lacZ plasmid. The STA1p-lacZ plasmid was transformed into FLO8-HA wild-type, FLO8-HA ste12Δ, FLO8-HA tec1Δ, and untagged strains. The resulting transformants were grown in synthetic medium containing 2% glycerol-ethanol to mid-log phase, and anti-HA (α-HA) ChIP assays were performed as described above with 1 mg of total extracts to determine Flo8 binding to UAS1-2. (C) Effect of Ste12 and Tec1 on lacZ expression from the endogenous STA1 promoter. The STA1p-lacZ plasmid was integrated into the original STA1 locus. The wild type and isogenic mutants were tested for β-galactosidase activity under the derepressed condition (2% glycerol-ethanol). (D) Binding of Flo8 to UAS1-2 of the endogenous STA1 promoter. FLO8-HA wild-type, FLO8-HA ste12Δ, FLO8-HA tec1Δ, and untagged strains were grown in synthetic medium containing 2% glycerol-ethanol to mid-log phase, and anti-HA ChIP assays were performed as described above with 1 mg of total extracts to determine Flo8 binding to UAS1-2 in the endogenous STA1 promoter.