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Fig. 4. Involvement of the C-terminal and of the coiled-coil domains in mitochondrial targeting. (A) Structure and localization (Loc.) of myc-tagged mitofusins and Mfn2-constructs. mt, mitochondria; ct, cytoplasma; ER, endoplasmic reticulum. (B-F) Cells were transfected with mitochondrial GFP (mtGFP) and/or with the indicated Mfn2-constructs. Myc-tagged Mfn2-mutants (myc) and the ER-marker calnexin were visualized with specific antibodies. (B) MF2-TMCT colocalizes with mtGFP in mitochondria. (C) MF2-IYFFT colocalizes with calnexin in the ER. (D-F) MF2-RRD, MF2-{Delta}C1 and MF2-{Delta}C2 are targeted to mitochondria, albeit inefficiently. A fraction of these molecules is mistargeted, especially at high expression levels. Bars, 15 µm.