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. 2005 Dec 6;392(Pt 3):537–544. doi: 10.1042/BJ20050806

Figure 1. Sal1p–yEGFP and endogenous Sal1p localize to mitochondria.

Figure 1

(A) Fluorescence microscopy of wild-type W303 cells expressing Sal1p–yEGFP (yeast Sal1p carrier fused to yEGFP from the pYeDP-SAL1-yEGFP vector) or yEGFP (from pYeDP-yEGFP vector) during exponential growth in 2% galactose. MitoTracker Red was used to locate mitochondria in the cells expressing Sal1p–yEGFP (MitoTracker), and phase-contrast microscopy was used to monitor the integrity of the cells. The same cells were visualized first with the MitoTracker filter set and then with a GFP filter set. Identical fields are presented. Scale bar, 5 μm. BF, bright field. (B) Immunoblot of subcellular fractions of wild-type W303 cells expressing Sal1p–yEGFP (left-hand panel) or yEGFP (right-hand panel) during exponential growth on 2% galactose. Blots were probed with antibodies against GFP and porin. M, mitochondria; PM, post-mitochondrial supernatant. Molecular-mass sizes are given in kDa. (C) Endogenous Sal1p was detected in the mitochondrial fraction. Subcellular fractions of wild-type W303 cells during exponential growth on 2% galactose were analysed by Western blotting with antibodies against Sal1p, Porin, P0, CPY and Vps10p. H, total homogenate; M, mitochondria; PM, post-mitochondrial supernatant.