Annexin-V staining to measure apoptotic cell death was performed as previously described [1 (link)]. The culture cells (1 × 105 cells/well) in the absence or presence of UA were collected into fresh tubes, and the cells were then washed with cold PBS and centrifuged at 2000 rpm for 2 min at room temperature. The cell pellets were mixed and reacted with 100 μL of Muse™ Annexin V and Dead Cell kit reagents (Millipore, Burlington, MA, USA) for 20 min. The apoptotic cells population was measured by using Mini Flow Cytometry Muse™ Cell Analyzer (Millipore, Burlington, MA, USA).
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