Interaction between Lte1 and activated Bud1. Cells coexpressing GST (MGY562) or GST-Lte1 (MGY563) and Bud1^G12V-HA under GAL1 promoter control were cultivated for 4 h in rich medium containing 2% galactose, HU was added, and cells were cultivated for another 2.5 h. After GST pull-down, bound proteins were eluted with 20 mM reduced glutathione. Total crude extract (CE) and eluted material (GST pull-down) were blotted and stained with α-HA antibody (top panels) or α-GST (bottom panel).