Mitochondrial Membrane Potential Assay

The JC-1 dye (MitoProbe JC-1 assay kit) was used to study the involvement of mitochondria during the early stages of apoptosis [23]. Jurkat cells were seeded in 24-well plates at a density of 3x10⁴ cells in 1 mL of media per well. Cells treated with 1 μM of 1, 50 μM carbonyl cyanide 3-chlorophenylhydrazone (CCCP), 1% v/v DMSO, and untreated cells were incubated for 7 h. At the end of the incubation period, cells were transferred to flow cytometry tubes and 1.5μL (200 μM) of JC-1 stock solution were added to each tube. After 30 min incubation at 37ºC, the JC-1 stained cells were analyzed via flow cytometry (Cytomics FC 500 Series Beckman Coulter).