FIGURE 6.

Singly anchored GRASP65 is binding-competent. Cells co-transfected with the indicated myc and GFP-tagged mitochondrial GRASP65 constructs were subjected to immunoprecipitation with anti-myc antibodies followed by immunoblotting with anti GRASP65 (G65, to detect myc-tagged constructs) or anti-GFP antibodies (to detect co-precipitation). Constructs were T20-G65 with either a myc or GFP tag followed by either a C-terminal anchor (ActA) or a stop codon (Stop). Co-transfection with an empty vector was used as a negative control (Empty). Loading control is 10% of total extract incubated with beads. Binding was quantified as the ratio of the amount of co-precipitated GFP-tagged protein to recovered myc-tagged protein (n = 3, ±S.E. (error bars)).