Quantitation of cell death was analyzed using the annexin V apoptosis assay on a Guava easyCyte flow cytometer (Millipore, Billerica, MA, USA), as previously described (11). Samples were prepared per the manufacturer’s instructions. Cells were classified into four categories based on dye uptake: viable cells (negative for annexin V and PI), early apoptotic cells (positive for annexin V only), late apoptotic cells (positive for annexin V and PI), and necrotic cells (positive for PI only).

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