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Fig. 6. Trafficking of Btn1p to the vacuole is Ypt7 dependent. (A) Localisation of GFP-Btn1 (green, left panel) in cells deleted for ypt7 and pre-labelled with FM4-64 (red, middle panel). GFP-Btn1 does not traffic to the vacuole in ypt7{Delta} cells. Bar, 10 µm. (B) Cells deleted for ypt7 or for ypt7 and btn1 were incubated with FM4-64 for 2 hours (left panel) then submitted to hypotonic shock (right panel). Vacuoles of ypt7{Delta} cells were smaller than those of cells deleted for ypt7 and btn1, and vacuoles of both strains were unable to fuse in response to hypotonic shock. Bar, 10 µm. (C) Vacuole size analysis in wild-type (wt) and cells deleted for btn1, ypt7 or ypt7 and btn1. Absence of Btn1p in ypt7{Delta} cells resulted in larger vacuoles. (D) Cells were incubated with CDCFDA for 10 minutes, to indicate relative intravacuolar pH. Cells deleted for ypt7 and btn1 had reduced CDCFDA fluorescence, indicating vacuoles were more alkaline than those of cells deleted for ypt7. Bar, 10 µm.





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