(A) Cultures of gle1-4, sup45-2, gle1-4 sup45-2, and wild-type control (WT) strains were serially diluted, spotted on YPD, and incubated 2–3 days at 23°or 30°C.
(B) Immunoprecipitations were performed with lysates from SUP35-TAP, SUP45-TAP, and wild-type (WT) yeast strains. Lysates (10% of input) and total immunoprecipitates (IP) were immunoblotted with α-mouse IgG (to detect TAP-tagged proteins (ProtA) or α-Gle1.
(C) Soluble binding assays were performed with GST-Sup45 or GST from bacterial lysates and recombinant purified Gle1. Gle1 input (10%) and bound samples were immunoblotted with α-GST or α-Gle1.