Btn1p localises upstream of Vps10p. (A,B) Btn1p localises to the Golgi
compartment in wild-type cells, which is upstream of Vps10p. Fluorescence
images of wild-type cells expressing GFP-Btn1p (A) or Vps10p-YFP (B) treated
with Brefeldin A (BFA) or without (DMSO control) showing that Btn1p-containing
Golgi compartments collapse back to the ER whereas Vps10p-containing
compartments do not. (C) Most Vps10p is not located at the Golgi. Confocal
images showing wild-type cells expressing Gms1p-CFP and Vps10p-YFP that do not
colocalise. (D) Bar chart depicting percentage of compartments
(n>200) containing indicated proteins. (E,F) Unlike Vps10p, Btn1p
remains Golgi localised in the absence of a functional retromer complex, and
is not misrouted to the vacuole. GFP-Btn1p (E) or Vps10p-YFP (F) expressed in
vps26Δ cells and treated with BFA or without (DMSO control).
All Btn1p-containing Golgi compartments collapse back to the ER whereas
Vps10p-containing vacuole compartments do not. Scale bars: 10 μm.