ChIP-seq of Mediator subunits in Mediator mutants and wild type yeast. ChIP-seq of Mediator subunits in Mediator mutants and wild type yeast

Mediator subunits Med17 (Srb4) and Med15 (Gal11) were subjected to ChIP in wild type yeast, med18∆ (srb5∆), med20∆ (srb2∆), and med2∆ med3∆ med15∆ (triple mutant) yeast, all in kin28-anchor away yeast after 1 hr rapamycin treatment to evict Kin28 from the nucleus. This results in inactivation of Kin28, which reduces Mediator turnover at promoters and allows stronger Mediator ChIP signal at promoters. Both Med17 and Med15 were tagged with myc and precipitated using the monoclonal antibody against c-myc. Please note that a specific control for rapamycin treatment was not performed in this experiment, as that has been well documented in the literature for this experiment, using what is called the “anchor away” method. The untagged sample is a control for non-specific immunoprecipitation.