Table 2. Intracellular localization of Bid-EYFP, tBid-EYFP and its deletion mutants.

Construct	EYFP/TRME	Localization
tBid	1±0,07	Mitochondrial
tBidE94G	1±0,07	Mitochondrial
tBidΔBH3	1±0,07	Mitochondrial
tBidΔH6	0,53±0,05	Cytoplasmic
tBidΔH6–H7	0,30±0,03	Cytoplasmic
tBidΔH6–8	0,27±0,03	Cytoplasmic
tBidΔBH3ΔH6–8	0,27±0,03	Cytoplasmic
tBidKK157,158AA	0,94±0,40	Mitochondrial*
tBidE94G,KK157,158AA	0,94±0,54	Mitochondrial*
tBidΔH3, or ΔH4, or ΔH5, or ΔH7, or ΔH8	0,97±0,09	Mitochondrial

CV-1 cells were transfected with plasmids encoding Bid-EYFP, tBid-EYFP or tBid-EYFP mutants in presence of 10 µM of Bok-D. 24 h later, cells were stained with 20 nM of the mitochondrial potential probe TMRE and the localization of the tBid-EYFP mutants was determined by calculation of the TMRE/EYFP ratios using microspectrofluorometry. A ratio of 1.0 indicates that EYFP exclusively colocalizes with TMRE at the mitochondria while a lower ratio indicates that EYFP is also localized to the cytosol. The * highlights the contructs that localize to the mitochondria but are unable to induce cytochrome c release or to alter mitochondrial bioenergetics. The constructs that localize to the cytoplasm are in italic.