Fig 3.

Cks1 is not required for the recruitment of Rtf1 to the GAL1 ORF. (A) Wild-type or cks1 deletion mutant cells were cultured in raffinose for 2 h, and 2% galactose was added for 45 min. GAL1 transcripts were quantified and normalized to actin (ACT1) mRNA. (B) Expression level of Flag-tagged Rtf1 in wild-type and cks1 deletion mutant strains. Samples were analyzed by SDS-PAGE and Western blotting. Amido black staining (bottom) was used as a loading control. (C) Recruitment of Flag-tagged Rtf1 to the GAL1 ORF in the absence of Cks1. Before and after galactose induction for 45 min, chromatin immunoprecipitation using anti-Flag antibodies was performed on wild-type untagged (control) wild-type Rtf1-Flag and cks1 Rtf1-Flag strains. Rtf1-associated chromatin fragments were isolated, amplified using the indicated primers, and normalized to the amount of input DNA prior to immunoprecipitation. Error bars represent SD.