Fig. 4.

MitoNEET is an integral membrane protein oriented toward the cytoplasmic face of the OMM. (A) Immunoblot analysis of swollen rat liver mitochondria treated to remove peripheral membrane proteins. Mitochondria were washed with either 200 mM KCl (high salt) or 0.1 M Na₂CO₃, pH 11.5 (high pH), and the proteins remaining in the pellets were separated by SDS/PAGE and immunoblotted. (B) SDS/PAGE and immunoblot analysis of trypsin-treated rat heart mitochondria (osmotically shocked). (C) SDS/PAGE and immunoblot analysis of trypsin-treated rat heart mitochondria (low pH washed). BCL2, B cell lymphoma-2; SMAC, second mitochondrial activator of caspases; VDAC, voltage-dependent anion channel protein.