Fitc annexin v 7 aad kit

The percentage of cells undergoing apoptosis and necrosis after treatment with different concentrations of methanolic root extract was quantified using FITC Annexin V-7-AAD kit (BD Pharmingen). HepG2 cells were seeded at a density of 2 × 10⁵ cells/well in 12-well plates and treated with different concentrations of extract (50, 100, and 200 μg/mL) for 24 h. The cells were harvested and resuspended in binding buffer and finally 5 μL of Annexin V-FITC and 5 μL of 7-AAD were added. Each tube was incubated in the dark for 15 min at room temperature. The stained cells were analysed on a FACS Canto II flow cytometer.

MOLT-16, LOUCY, and ALL-SIL cells were treated with DMSO and the EC₅₀ (nM) of venetoclax, volasertib, and a combination in 6-well plates for 24, 48, and 72 h. Then, JURKAT, DND-41, CML-T1, TALL-1, and RPMI-8402 were treated in 6-well plates for 72 h with DMSO, venetoclax (50, 100, and 500 nM), and the respective concentrations of volasertib for each cell line along with the combination treatment. After each incubation period, the cells were processed, and the apoptotic cells were quantified using the FITC-Annexin-V/7-AAD kit (BD Biosciences, USA) according to the manufacturer’s protocol. Finally, the data were analyzed with the FlowJo software.