A549 cells were seeded in a six-well plate at a concentration of 1.0 × 105 cells/mL, incubated for 24 h at 37 °C in a humidified atmosphere of 5% CO2, and then were treated with DDP (50 μM) and FXS-3 at different concentrations (5.4, 16.8, and 50 μM). After an incubation of 48 h, A549 cells were collected, washed with phosphate-buffered saline (PBS), and then fixed with cold 70% ethanol for at least 24 h. Then, A549 cells were washed twice with PBS, stained with PI/RNase staining solution (Sigma-Aldrich, St. Louis, MO, USA) for 30 min in the dark, and then analyzed using a FACSCalibur flow cytometer (BD INSTRUMENTS INC., San Jose, CA, USA).
Pi rnase staining solution
Manufactured by Merck Group
Sourced in United States
The PI/RNase staining solution is a laboratory reagent used for the detection and quantification of cellular DNA content. It contains propidium iodide, a fluorescent dye that binds to DNA, and RNase, an enzyme that degrades cellular RNA. This solution is commonly used in flow cytometry analysis to measure the DNA content of individual cells, which can provide information about cell cycle distribution and ploidy.
Automatically generated - may contain errors
3 protocols using pi rnase staining solution
1
Cell Cycle Analysis of A549 Cells
2
Cell Cycle Analysis by Flow Cytometry
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Cell cycle distribution was determined using propidium iodide (PI) staining and flow cytometry analysis. Briefly, HT-29 cells (2×106 cells/dish) were treated with 65 μg/mL JCo extract for 0, 6, 12, 24, and 48 h, followed by the addition of PI/RNase staining solution (40 μg/mL of PI and 100 μg/mL of RNase; Sigma) and incubation at 4°C overnight. The DNA content (FL2 intensity) of each cell was measured using FACScan (Beckton Dickinson, USA) and Kaluza Flow Cytometry Analysis software (version 1.2, Beckman Coulter, USA).
3
Cell Cycle Analysis of Turmesac® and Camptothecin
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Cells were cultured at a density of 2 × 10 5 cells/2 ml and incubated at 37°C for 24 h. Cells were replenished with fresh media and treated with Turmesac ® or camptothecin. Cells were harvested and centrifuged for 5 min at 300 × g. Cells were rinsed with PBS and fixed in 1 ml of cold 70% ethanol on ice for 30 min. Cells were centrifuged and washed twice with PBS. After the wash step, cells were stained with 400 µl PI/RNAse staining solution (Cat No: 550825, Sigma) and incubated for 20 min at room temperature. Using the BD FACSCalibur flow cytometer, cells were analyzed for cell cycle arrested.
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