Cell quest research software

Jurkat cells (1.0 × 10⁵ cells/tube) were incubated
in 10% FBS RPMI 1640 medium containing solution of 1 (10⁵ μM) and cisplatin (0–50 μM) for 24 h.
The cells were then centrifuged at 3000 rpm for 5 min at 4 °C,
then the supernatant was discarded, and the pellet was resuspended
in 200 μL of PBS. The cells were then centrifuged at 3000 rpm
for 5 min at 4 °C, then the supernatant was discarded, and the
pellet was resuspended in 195 μL of 1 × binding buffer.
A 195 μL aliquot of the sample solution was incubated with 5
μL of FITC-conjugated annexin V (Invitrogen) for 15 min at room
temperature in the dark. Then the cells were then washed by 1 ×
binding buffer and added 190 μL of 1 × binding buffer 10
μL of PI (20 μg/mL, Invitrogen). 200 μL of the sample
solution transferred to culture tube were analyzed by flow cytometry
(Becton Dickinson) using Cell Quest Research Software (Becton Dickinson).

Annexin V-FITC/PI stained fluorescence-activated cell sorter (FACS) and Annexin V-FITC stained fluorescence microscopy were used to measure cell apoptosis. Briefly, A549 cell lines were seeded at a density of 2 × 10⁴ cells/well overnight, divided into different groups (n = 3), and then treated with GJXLT at different concentrations for 24 h. All cells were harvested through trypsinization and washed twice with cold PBS (0.15 mol/L, pH 7.2). The cells were centrifuged at 1000 r/min for 5 min. Then, the supernatant was discarded and the pellet was resuspended in 1× binding buffer at a density of 1.0 × l0⁶ cells/mL. A total of 100 μL of the sample solution was transferred to a 5 mL culture tube and incubated with 5 μL of FITC-conjugated Annexin V and 10 μL of PI for 15 min at room temperature in the dark. A total of 400 μL of 1× binding buffer was added to each sample tube, and the samples were analyzed by FACS (Becton Dickinson, USA) using Cell Quest Research Software (Becton Dickinson, USA).