Flow cytometry analysis was conducted as described above [12 , 13 (link)]. Cells (8 × 103) were seeded into 6-well plates and cultured for 24 h. After culture with DDP and Miao for 48 h, cells (1 × 105/ml) were collected and suspended in 1 mL binding buffer containing 10 µL Annexin V-FITC and 10 µL propidium iodide (PI) (Keygene Biotech, China). After incubation for 10 min in the dark, apoptosis was measured through flow cytometry. The apoptotic rate was scored by quantifying late apoptosis or necrosis cells (Annexin V-FITC+ PI+) and early apoptosis (Annexin V-FITC+ PI−).
Propidium iodide
Manufactured by Keygene
Sourced in China
Propidium iodide (PI) is a fluorescent dye used in molecular biology and flow cytometry applications. It is a nucleic acid intercalator that binds to DNA. PI can be used to stain and identify cells with compromised cell membranes, which is commonly associated with cell death.
Automatically generated - may contain errors
4 protocols using propidium iodide
1
Flow Cytometry Analysis of Apoptosis
2
Apoptosis Assay via Flow Cytometry
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After cell transfection, the supernatant and cells were collected through centrifugation. The precipitates were suspended in 600 μL binding buffer and added with 5 μL Annexin V-FITC (KeyGene Biotech, China). After mixing, 5 μL propidium Iodide (PI) (KeyGene Biotech, China) was added, and the reaction was conducted at room temperature in a dark environment for 15 min. Flow cytometry was used to detect the percentage of cell apoptosis.
3
Apoptosis Quantification by Flow Cytometry
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Cell apoptosis was determined through Flow cytometry. The cells were digested by trypsin and collected. Following resuspension in 500 μl binding buffer, the cells were added with 5 μl Annexin V-FITC and 5 μl Propidium Iodide (PI) (KeyGene, Nanjing, China) at room temperature in the dark for 15 min. The cells were subsequently tested by flow cytometry.
4
Cell Cycle and Apoptosis Analysis
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For cell cycle assay, A549/Taxol and H1299/Taxol cells were collected after transection and fixed in ethanol (70%; Beyotime) for 12 h at −20℃. After fixation, the cells were harvested via centrifugation, followed by addition of propidium iodide (PI; KeyGene) and RNase A (Beyotime). FACScan® flow cytometry was employed to detect cell cycle distribution.
For detection of cell apoptosis, the transfected cells were harvested. After suspending in binding buffer (0.5 ml), cells were labeled with Annexin V‐FITC and PI (Beyotime), followed by measurement of apoptotic cells with FACScan® flow cytometry.
For detection of cell apoptosis, the transfected cells were harvested. After suspending in binding buffer (0.5 ml), cells were labeled with Annexin V‐FITC and PI (Beyotime), followed by measurement of apoptotic cells with FACScan® flow cytometry.
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