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Figure 1


Fig. 1. Identification of the Mfn-binding protein (MIB). (A) Isolation of MIB using MBP-Mfn{Delta}C-conjugated resin from rat liver cytosol. MBP-Mfn1{Delta}C- or MBP-conjugated resin were treated with the indicated concentrations of NaCl. The eluted proteins were analyzed by SDS-PAGE and CBB-staining. (B) Schematic representation and sequence identity of medium-chain dehydrogenase/reductase protein superfamily. dADH, medium-chain alcohol dehydrogenase; QOR, E. coli quinone oxidoreductase. (C) Hydropathy profile of rat MIB deduced by Kyte and Doolittle algorithm with a 10-amino-acid-residue window. (D) Elution profile of MIB-FLAG from MBP-Mfn{Delta}C-conjugated resin. Cytosolic fraction prepared from MIB-FLAG-expressing HeLa cells was incubated with MBP-Mfn1{Delta}C beads, then eluted by buffer containing the indicated concentrations of NaCl as in (A). The eluted proteins were analyzed by SDS-PAGE and subsequent immunoblotting using anti-flag antibody.





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