Tda2 localizes to sites of CME at late stages of the endocytic process and independently of the clathrin coat. (A) Live-cell fluorescence microscopy showing strong colocalization between Tda2-GFP and Sla1-RFP. (B) Dynamics of Tda2-GFP and Sla1-RFP were compared. (Left) One frame of a video indicating with a white box the endocytic site used for constructing a kymograph. (Right) Kymograph illustrating how Tda2 is recruited to late stages of endocytosis. (C) Live-cell fluorescence microscopy showing strong colocalization between Tda2-mCherry and Rvs167-GFP. Notice Tda2-mCherry arrives a few seconds before Rvs167-GFP. (D and E) Patch lifetimes of Sla1-RFP and Tda2-GFP were measured in WT cells (CHC1) and cells carrying a temperature-sensitive allele of the clathrin heavy chain (chc1-ts) after incubating the cells at 23° or 37°. Clathrin disruption (37°) reduces patch lifetime in Sla1-RFP but not Tda2-GFP. Bars, 1 µm. Error bars indicate means ± SEM. *, P < 0.05.