Figure 6.

Overexpression of Tul1 causes a constitutive degradation of a subset of vacuolar membrane proteins. (A) Localization of chromosomally tagged Ypq2-GFP, Fth1-GFP, and Vph1-GFP in cells overexpressing Ssh4, Tul1, or the empty vector. (B) Western blot analysis of protein levels for chromosomally tagged Ypq2-GFP, Fth1-GFP, and Vph1-GFP in cells overexpressing Ssh4 (S-OE), Tu1(T-OE), or the empty vector (Vec). G6PDH was used as a loading control. 1 OD₆₀₀ of cells were loaded for Ypq2 and Fth1 samples, 0.4 OD₆₀₀ of cells were loaded for Vph1 samples owing to the high expression level of Vph1-GFP. (C) Localization of chromosomally tagged Ypq1-GFP, Zrt3*-GFP, Zrc1-GFP, and Cot1-GFP in cells overexpressing Ssh4, Tul1, or the empty vector. (D) Western blot analysis of protein levels for chromosomally tagged Ypq1-GFP, Zrt3*-GFP, Zrc1-GFP, and Cot1-GFP in cells overexpressing Ssh4, Tul1, or only the empty vector. G6PDH was used as a loading control. 1 OD₆₀₀ of cells were loaded for each lane. (E) Localization of chromosomally tagged Ycf1-GFP, Vba4-GFP, and Ypl162c-GFP in cells overexpressing Ssh4, Tu1, or the empty vector. (F) Western blot analysis of protein levels for chromosomally tagged Ycf1-GFP, Vba4-GFP, and Ypl162c-GFP in cells overexpressing Ssh4, Tul1, or the empty vector. G6PDH was used as a loading control. 1 OD₆₀₀ cells were loaded for each lane. Bar, 1 µm.