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. 2014 Oct 28;111(45):16017–16022. doi: 10.1073/pnas.1408061111

Fig. 4.

Fig. 4.

In vitro Opa1 C-cleavage is executed by a cysteine protease that acts downstream of Oma1 but that requires Mfn2. (A and B) In vitro Opa1 C-cleavage assay generates Opa1 fragments that have the same electrophoretic mobility of endogenous CTF-1 and CTF-2. An asterisk denotes an unspecific product of the in vitro Opa1 C-cleavage. (C) Opa1 C-cleavage is blocked by the cysteine protease inhibitor E-64. (D and E) Silencing the mitochondrial cysteine protease CAPN1 and genetic ablation of the Opa1 protease Oma1 do not affect Opa1 C-cleavage in vitro. (F) Genetic ablation of Mfn2, but not of Mfn1, blocks Opa1 C-cleavage. Mitochondria were prepared from Mfn1−/− Mfn2−/− MEFS and incubated for the indicated time with DDM to activate Opa1 C-cleavage and the generation of CTF-1 and CTF-2.