Figure 5

Parkin affect the stability of the MICUs monomers and MICU1/MICU2 heterodimers. HeLa cells were co-transfected with MICU1-HA, MICU2-FLAG and the indicated Parkin constructs. 24 hours later cells were either untreated or treated with 10 μM CCCP for 2 hours (where indicated). Cells were then harvested, lysed and subjected to western blot analysis with the indicated antibodies. (A) Representative western blot showing the half-life of co-expressed MICU1 and MICU2, with or without CCCP treatment. Densitometry analysis of MICU1 (B) and MICU2 (C) protein levels, normalized to endogenous MCU protein, in cells co-expressing MICU1 and MICU2. (D) Non-reducing western blot analysis and (E) quantification of the MICU1/MICU2 heterodimers in the presence of the indicated Parkin constructs. Each bar represents mean +/− S.E.M of at least three independent experiments; *p < 0.05; **p < 0.01. a.u.: arbitrary units.