Mitochondrial membrane potential (Δψm) was measured by flow cytometry using a fluorescent cationic dye, DePsipher, according to the vendor’s protocol (DePsipher, R&D System Inc.), as described previously [19 (link),20 (link),21 (link)]. The dye readily enters healthy mitochondria and fluoresces red in its multimeric form. However, in apoptotic cells, the mitochondrial membrane potential collapses, and the dye cannot enter the mitochondria and remains in the cytoplasm in its green fluorescent monomeric form.
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