FIGURE 4:
Coiled coils contribute to endocytic-site localization by aggregating Ede1 molecules. (A) Representative images of Ede1-GFP or ede1ΔCCinternal-GFP tagged with a prenylation motif (CCIIS) or a control sequence (SSIIS). Scale bar, 5 μm. (B) Constructs from A were expressed with Sla1-mCherry and imaged for 4 min at 2 s/frame with ∼300-ms delay between channels. The first frame of a movie of a representative cell, followed by a circular kymograph of the bud and mother cortex. Scale bars, 2 μm and 40 s. (C) Whole-cell lysates of diploid cells with the indicated genotype were subjected to immunoadsorption using mouse anti-GFP antibody, followed by SDS–PAGE and Western blotting, using rabbit anti-GFP antibody to detect Ede1-GFP and mouse anti-myc antibody to detect the Ede1-13xmyc-tagged truncations.