SKOV3/PTX and HeyA-8/PTX cells were seeded into six-well plates and treated with PTX for 48 h. To assess the cell cycle and apoptosis, 3 × 105 treated cells were seeded into six-well plates and cultured for 48 h at 37°C. The cells for cell cycle analysis were digested using trypsin (HyClone), washed twice with phosphate-buffered saline (PBS), and fixed in 70% ethanol overnight at 4°C. The cells were centrifuged at 500 × g for 5 min, washed twice with cold PBS, and centrifuged. After treating with RNase A (0.1 mg/mL) and propidium iodide (PI, 0.05 mg/mL) purchased from 4A Biotech (Beijing, China) for 30 min at 37°C, cell cycle analysis was performed through fluorescence-activated cell sorting flow cytometry (Beckman Coulter, Palo Alto, CA, USA). For the analysis of apoptosis, cells were trypsinized followed by two PBS washing steps. The cells were stained using the annexin V/PI detection kit (4A Biotech, Beijing, China) for 5 min at room temperature. The apoptotic cells were measured using flow cytometry (Beckman Coulter). All experiments were repeated at least three times.
Propidium iodide pi
Manufactured by 4A Biotech
Sourced in China
Propidium iodide (PI) is a fluorescent dye commonly used in flow cytometry applications. It is a nucleic acid intercalator that selectively binds to DNA, emitting a red fluorescence upon excitation. PI is often used to determine cell viability and assess cell cycle status.
Automatically generated - may contain errors
2 protocols using propidium iodide pi
1
Cell Cycle and Apoptosis Analysis
2
Cell Cycle and Apoptosis Analysis
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SKOV3/PTX and HeyA-8/PTX cells were seeded into 6-well plates and treated with PTX for 48 h. To assess the cell cycle and apoptosis, we seeded 3 × 105 treated cells into 6-well plates and cultured them for 48 h at 37°C. The cells for cell-cycle analysis were digested using trypsin (Hyclone), washed twice with phosphate-buffered saline (PBS), and fixed in 70% ethanol overnight at 4°C. The cells were centrifuged at 500 × g for 5 min, washed twice with cold PBS, and centrifuged. After treatment with RNase A (0.1 mg/mL) and propidium iodide (PI, 0.05 mg/mL) purchased from 4A Biotech (Beijing, China) for 30 min at 37°C, cell-cycle analysis was performed through fluorescence-activated cell sorting flow cytometry (Beckman Coulter, Palo Alto, CA, USA). For the analysis of apoptosis, cells were trypsinized followed by two PBS washing steps. The cells were stained using the Annexin V/PI detection kit (4A Biotech, Beijing, China) for 5 min at room temperature. The apoptotic cells were measured using flow cytometry (Beckman Coulter). All experiments were repeated at least three times.
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