Nob1 Overexpression in Gal Cells

Gal::Nob1 cells grown in glucose media were supplemented with either WT Nob1 or an empty vector. Cells were harvested in mid-log phase, and reporter assays were carried out essentially as described before [6 (link)]. Cells were lysed, and luciferase activity was measured with the Promega Dual-Luciferase Reporter Assay System on a PerkinElmer EnVision 2104 Multilabel Reader according to the manufacturer’s protocol, with assay volumes scaled down to 15%. For each sample, firefly luciferase activity was normalized against renilla activity; subsequently, values observed for depleted Nob1 were normalized against those for WT Nob1.