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Figure 3


Fig. 3. Bem1p can interact with the exocyst complex through Sec15p. (A) Purified recombinant GST-Bem1p (GST-Bem1) and GST were resolved by SDS-PAGE and the gel was stained with Coomassie Brilliant Blue. Proteins of the predicted molecular masses were observed. (B) Exocyst complex purified from yeast strain NY2521 was resolved by SDS-PAGE and the gel was silver stained to visualize the subunits. (C) GST-Bem1p (GST-Bem1) or GST immobilized on glutathione-Sepharose beads was incubated with the purified exocyst complex as described in the Materials and Methods. Lanes 1 and 2 represent 2.5% and 5%, respectively, of the exocyst complex used in the binding reactions. The beads were washed four times with the binding buffer and the bound proteins were eluted with SDS-PAGE sample buffer. The eluates were then resolved by SDS-PAGE, and western blot analysis was performed using anti-HA (to detect Sec5-3xHA), anti-Sec15 and Sec10 antibodies.





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