Annexin 5 fitc propidium iodide apoptosis kit

Cell apoptosis was determined using an Annexin V-FITC/propidium iodide (PI) apoptosis kit (BD, San Jose, CA, USA) by flow cytometry. Treated cells were pelleted by centrifugation at 800 × g for 5 min, washed twice with cold PBS, and resuspended into 400 μl binding buffer. Ten microliters of Annexin V-fluorescein isothiocyanate (FITC) solution and subsequently 5 μL of PI per 100 μL of cell suspension were then added, and the mixture was incubated for 15 min in the dark at room temperature (20-25°C). Finally, the fluorescence of the cells was recorded using the Accuri C6 flow cytometer (BD, USA).

Annexin V-FITC/propidium iodide (PI) apoptosis kit (BD Biosciences, CA, USA) was applied to evaluate cell apoptosis. Transfected PAMSCs were trypsinized, harvested, and then collected after centrifugation. Cells were stained with Annexin V and PI according to the instruction after washed by PBS (cold). The early as well as late apoptotic cells were detected using flow cytometry and Cell Quest Pro software (BD Bioscience).

Flow cytometry was performed using previously described methods [22 (link), 25 (link)]. HN6 and Cal27 cells transfected with si-lincRNA-p21, lincRNA-p21, or scramble were harvested 48 h after transfection. After incubating with reagents from the Annexin V-FITC/propidium iodide (PI) apoptosis kit (BD Biosciences, Franklin Lakes, NJ, USA), cells were analysed using a BD FORTASA flow cytometer (BD Biosciences) and the FlowJo software. For cell cycle analysis, cells were incubated using the PI/RNase staining kit (BD Biosciences). The subsequent steps were performed as described above.

Annexin V-FITC/propidium iodide (PI) Apoptosis kit (BD Biosciences, USA) was used to measure the apoptosis level of A375 cells treated with IC50 concentration of ACY-1215, tubastatin A, and EHT 1864, respectively. The cells were stained with Annexin V and PI according to the instructions and then analyzed by flow cytometry (Beckman, USA). Early apoptotic A375 cells showed annexin V-positive and PI-negative, and late apoptotic A375 cells were stained with both annexin V- and PI-positive. The results were presented as the percentage of apoptotic cells.

MCF‐7 cells were seeded in 6–well plates at a density of 1 × 10⁶ cells per well, grown for 24 h before treatment with nanocomposites at IC50 value for 48 h. The apoptosis was examined by flow cytometry using the Annexin V‐FITC/propidium iodide (PI) apoptosis kit (BD Biosciences), according to the manufacturer's instruction. Briefly, MCF‐7 cells were washed with cold PBS and were then resuspended in 1 mL of 1x binding buffer (provided with kit) at density of 10 × 10⁶ cells/mL. 100 µL of cell suspension was incubated with 5 µL of Annexin V‐FITC and PI for 20 min in the dark at room temperature, and then examined by flow cytometry. The percentage of cells in different stages (apoptosis and necrosis) was examined by FlowJo 7.6 software. At least 12,000 events were recorded and represented as dot plots.