Fig. 5.

Mfn2 ablation impairs mitochondrial Ca²⁺ uptake in an MCU-independent manner. (A and B) Equal amounts (40 μg) of total lysates from cells of the indicated genotypes were separated by SDS/PAGE and immunoblotted using the indicated antibodies. (C) Mean ± SEM of densitometric quantification of levels of the indicated MCU holoplex components from 3 to 10 independent experiments as in A and B. (D) MEFs of the indicated genotype were infected where indicated with MCU expressing adenoviruses and, after 48 h, were lysed and equal amount of proteins (40 µg) separated by SDS/PAGE and immunoblotted with the indicated antibodies. (E) Recordings of [Ca²⁺]_i in Ca²⁺-free KRB in cells of the indicated genotypes overexpressing MCU. Where indicated, MEFs were perfused with 0.2mM (WT) or 10 µM (Mfn2^−/−) ATP. (F) Mean ± SEM of three independent experiments of peak [Ca²⁺]_i from experiments performed as in E. (G) Recordings of [Ca²⁺]_mit in experiments performed as in E. (H) Mean ± SEM of peak of [Ca²⁺]_mit from three independent experiments performed as in G. *P < 0.05 vs. WT.