Quantifying Mitochondrial Co-localization

Immunofluorescence confocal microscopy was carried out as described^{13 (link), 19 (link)}. For mitochondrial staining, the MitoTracker Red CMXRos (500 nM, Molecular Probes) was added to cultures 15 min before fixation. Specimens immunostained with different antibodies were examined in the SP5 confocal microscopy system (Leica). Quantitative co-localization analysis of confocal images was performed with the Pearson’s correlation coefficient (r) using the Leica integrated program. The Pearson’s correlation coefficient was presented as mean ± SEM by the Student’s t-test software (http://www.physics.csbsju.edu/stats/t-test.html). Measurement of immunofluorescence intensity was performed by the Leica confocal microscopy software.