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. 2011 Mar 8;6(3):e16746. doi: 10.1371/journal.pone.0016746

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Rakovic et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Fibroblasts from (A) a healthy control, (B) a homozygous PINK1 mutant and (C) a homozygous Parkin mutant were treated with 1 µM valinomycin alone (left panel) or with 1 µM valinomycin plus 10 µM epoxomicin (right panel). Proteins were extracted at different time points and analyzed by Western blotting. In control cells, valinomycin treatment initiated the ubiquitination of Mfn2 after 1 h of incubation. This effect was prevented by simultaneous exposure to epoxomicin. The mitochondrial marker VDAC1 and the cytosolic marker β-actin served as loading controls. Mfn2 – mitofusin 2; Ub-Mfn2 – ubiquitinated mitofusin 2; VDAC1 – voltage-dependent anion channel 1.