Quantifying Mitochondrial Membrane Potential

Was performed as described^{68 (link)}. For Flag detection, DYKDDDDK Tag Rabbit antibody was used at 1:100 dilution (Cell Signaling Technology, 14793). Where indicated, 30 min before fixation with 4% paraformaldehyde, cells were incubated with 100 nM MitoTracker Red CMXRos (Invitrogen, M7512) to assess mitochondrial membrane potential. Mounted coverslips were imaged on a Leica TCS SP8 Confocal Laser Scanning Microscope (Leica Microsystems) with ×63 objective and 1.4 numerical aperture. Quantification of MitoTracker Red CMXRos fluorescence intensity per cell was performed using ImageJ (NIH) and expressed as mean integrated density. For detection of BODIPY FL C₁₆ in vivo, sections from freshly isolated livers were mounted with Fluoromount-G medium (SouthernBiotech, 0100) and imaged on Leica TCS SP8 Confocal Laser Scanning Microscope with ×10 objective and 1.4 numerical aperture.