Fxcycletm pi rnase solution

For apoptosis analysis, 1.5 × 10⁵ cells/well were seeded in 6-well plates. After overnight incubation, the cells were treated with or without quercetin (50, 100, 200 μM) and luteolin (10, 20, 40 μM) for 48 h. The cells were collected and suspended in a 100 μl binding buffer containing 5 μl FITC-conjugated Annexin V and 5 μl PI (Annexin V FITC Apop Dtec, BD). After dark incubation for 15 min at room temperature, a 400 μl binding buffer was added. The apoptosis condition of the cell samples was analyzed using the Attune Flow cytometer (ThermoFisher, USA) within 1 h.
For cell cycle analysis, the cells were collected and fixed with 75% ethanol at −20°C overnight. Then, the cells were incubated with 0.5 ml propidium iodide (PI, 0.5 ml FxCycleTM PI/RNAse Solution, BD) in the dark for 45 min. The cell samples were tested using the Attune Flow cytometer (ThermoFisher, USA) for cell cycle analysis.

For apoptosis and cell cycle analyses, 1.5 × 10⁵ cells/well were seeded in 6-well plates. After overnight incubation, the cells were treated with or without CNC extract (25, 50, 100, 150 μg/mL) for 48 h. Cells were collected and performed according to the protocol of the Annexin V FITC Apoptosis kit (and FxCycleTM PI/RNAse Solution (both from BD Biosciences, Franklin Lakes, NJ, USA). All cell samples were analyzed on the Attune Flow cytometer (Thermo Fisher Scientific) within 1 h.