In vitro protein-binding assays testing the interaction network of Bcd1 with its binding partners.
A, MBP-Rsa1/Hit1 immobilized on amylose resin can pull down WT and D72A variant Bcd1. B–D, MBP-Bcd1 (WT) or MBP-Bcd1-D72A (D72A) bound to amylose resin pulldowns Rvb1/2 (B), Nop58 (C), and Tah1/Pih1 (D). E, GST-Snu13 immobilized on GSH Sepharose resin can pull down WT but fails to interact with Bcd1-D72A. M, molecular weight marker (kDa); I, input; W, last wash; E, eluate; *, nonspecific contaminant.