Bax linked to cyan fluorescent protein (CFP-Bax), and the marker of the OMM, 20 amino acids from the COOH terminus of Bax linked to yellow fluorescent protein (YFP-20) were described previously (Nechushtan et al., 1999 (link)). Mfn2-YFP was constructed from the GFP construct described previously (Santel and Fuller, 2001 (link)). Drp1 and Drp1K38A expression vectors were described previously (Smirnova et al., 1998 (link); Frank et al., 2001 (link)). Drp1 was recloned into pEYFP-C1 to yield pEYFP-Drp1 (YFP-Drp1).
Human homologues of Fis1 (Mozdy et al., 2000 (link)), OMP 25, and synaptojanin 2A (Nemoto and De Camilli, 1999 (link)) were cloned from fetal human brain mRNA (Stratagene) using RT-PCR, and sub-cloned into pEYFP-C1 vectors (BD Biosciences). Mito-YFP (BD Biosciences) and mito-DsRed2 (BD Biosciences) revealed mitochondria.
Cells were transfected using the FuGENE 6 (Roche) and treated as indicated in individual experiments, 12–15 h later.
Human homologues of Fis1 (Mozdy et al., 2000 (link)), OMP 25, and synaptojanin 2A (Nemoto and De Camilli, 1999 (link)) were cloned from fetal human brain mRNA (Stratagene) using RT-PCR, and sub-cloned into pEYFP-C1 vectors (BD Biosciences). Mito-YFP (BD Biosciences) and mito-DsRed2 (BD Biosciences) revealed mitochondria.
Cells were transfected using the FuGENE 6 (Roche) and treated as indicated in individual experiments, 12–15 h later.
