S3483 compound induces apoptosis in differentiated HL-60 cells and PMNs, and DPI and Trolox C prevent the apoptosis induction.

PMNs, differentiated and undifferentiated HL-60 cells, and Jurkat cells (2 × 10⁶) were incubated (at 37°C in 5% CO₂/95% air for 60 minutes) in the complete RPMI 1640 culture medium containing the solvent alone (10 μL/mL ethanol) or 100 μM S3483. Where indicated, 10 μM Trolox C or 10 μM DPI was also added to the cell suspension. Cells stained with the 2-integrin reagent VIM12 (to reveal phosphatidylserine exposure on cell surface) and propidium iodide were observed with a real-time confocal microscope. Apoptotic cells were identified on the basis of the phosphatidylserine exposure on the plasma membrane surface and on morphologic changes. A minimum of 200 cells was scored for each sample. Values are means ± SEMs of 3 to 5 independent experiments.