Annexin 5 pe 7 amino actinomycin 7 aad apoptosis detection kit

Cell apoptosis was analyzed using an Annexin V PE/7-Amino-Actinomycin (7-AAD) Apoptosis Detection Kit (BD Pharmingen, San Diego, CA, USA) according to the manufacturer’s protocol. The following groups were used to set up compensation and to define quadrants: unstained control cells, cells stained with PE Annexin V (no 7-AAD), and cells stained with 7-AAD (no PE Annexin V). Apoptotic cells were examined using a flow cytometer (Becton-Dickinson, USA) within 1 h, and the percentage of early apoptotic cells (upper-right quadrant) was measured using FlowJo software (Tree Star, Ashland, OR, USA).

As the manufacturer’s instructions for a Cell-Light EdU DNA cell proliferation kit (#C103010-1, RiboBio, China) described, transfected cell lines were seeded in 96-well plates and incubated with EdU in medium (50 μM) for 2 hours. Then, cells were washed twice and fixed in 4% paraformaldehyde for 10 min, permeabilized with 0.5% Triton-X 100, and stained with Apollo® fluorescent dye. Photographs of cells were independently taken with an OLYMPUS confocal microscope.
The cells were analyzed by FACS according to the standard protocol provided by the manufacturer (BD FACSAria II). Apoptosis was measured by using an Annexin V-PE/7-Amino-Actinomycin (7-AAD) Apoptosis Detection Kit (#559763, BD Biosciences Pharmingen, US). After treatment with 5-FU (#9648, TargetMol, China) at the indicated concentrations for 48 hours, cells were harvested by trypsinization, washed with cold PBS and then resuspended in 1X binding buffer. Then, 5 μl of PE Annexin V and 5 μl 7-AAD were added to each tube. The suspension was then mixed well and incubated for 15 min in the dark at room temperature (RT) (25°C). After resuspension, samples were analyzed by flow cytometry.

The cells were analyzed by FACS according to the standard protocol provided by the manufacturer (BD FACSAria II). Apoptosis was measured by using an Annexin V-PE/7-Amino-Actinomycin (7-AAD) Apoptosis Detection Kit (#559763, BD Biosciences Pharmingen, US). After treatment with 5-FU (#9648, TargetMol, China) at the indicated concentrations for 48 hours, the cells were harvested by trypsinization, washed with cold phosphate-buffered saline (PBS), and then resuspended in 1X binding buffer. Then, 5 µL of PE Annexin V and 5 µL of 7-AAD were added to each tube. The suspension was then mixed well and incubated for 15 min in the dark at room temperature (RT) (25°C). After resuspension, the samples were analyzed by flow cytometry.