Figure 6.

Deletion of Ysc84 affects the invagination stage of endocytosis. (A) Effect of deletion of ysc84 on markers of different stages of endocytosis was assessed. Lifetime of Sla1-GFP, Las17GFP, and Abp1GFP was assessed by measuring patch lifetime from movies recorded of cells expressing these markers. In all cases, there is a significant reduction in patch lifetime. More than 10 patches were assessed in wild-type and mutant strains. (B) The behavior of patches was also analyzed using kymographs with the marker Sla1GFP. As shown in the top panels in wild-type cells, Sla1GFP localizes to the plasma membrane and then shows a clear inward movement before disassembling. In the absence of ysc84 (bottom), the lifetime on the membrane and the extent of the inward movement is reduced. (C) To determine whether the Ysc84 homologue Lsb3 plays a distinct or overlapping role from Ysc84, the effect of deletion of lsb3 on the invagination step of endocytosis was also assessed. More than 15 patch lifetimes were measured for each strain. Differences between patch lifetimes in wild-type and Δysc84 and between Δysc84 and Δysc84Δlsb3 cells are significant (p value <0.001 and 0.0096, respectively).