Supplementary Figure 4: Electronic gating strategy for analysing cell death based only on TO-PRO-3.
From: Monitoring the progression of cell death and the disassembly of dying cells by flow cytometry
Flow cytometry analysis showing a one-stage electronic gating strategy used to identify viable cells, early apoptotic cells (A5− and A5+), necrotic cells, as well as apoptotic bodies and debris. Human Jurkat T cells were induced to undergo apoptosis by anti-Fas treatment (31.25 ng ml−1, 4 h). Samples were stained with A5-FITC (1 in 200 dilution), 7-AAD (1 μg ml−1) and TO-PRO-3 (0.5 μM) in 1x A5 Binding Buffer. 30,000 events (cells and cell fragments) were recorded by the FACSCanto II flow cytometer and resultant data analysed by the FlowJo 8.8.6 software. The A5-FITC and 7-AAD parameter was not used during data analysis.
