Cells were seeded in six-well plates at a density of 1 × 106 cells per well. Twelve hours later, circ-ITCH-overexpressing or circ-vector cells were transfected with miRNA mimics or miR-NC. After seeding for 48 h, all cells were collected and stained with Annexin-V–fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis assay kit (Yeasen, Shanghai, China), and then analyzed using a flow cytometry (Epics XL-MCL; Beckman Coulter, Brea, CA, USA). The experiments were repeated three times.
Annexin 5 fluorescein isothiocyanate fitc propidium iodide apoptosis assay kit
Manufactured by Yeasen
Sourced in China
The Annexin-V–fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis assay kit is a laboratory tool used to detect and quantify apoptosis in cell samples. It combines the use of Annexin-V, a protein that binds to phosphatidylserine, and propidium iodide, a DNA-binding dye, to identify cells in various stages of apoptosis and necrosis.
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2 protocols using annexin 5 fluorescein isothiocyanate fitc propidium iodide apoptosis assay kit
1
Apoptosis Evaluation of Circ-ITCH Overexpression
2
Detecting HTR-8/SVneo Cell Apoptosis
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In compliance with an existing protocol [15 (link)], flow cytometry was conducted to detect HTR-8/SVneo apoptosis. The Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis assay kit (YEASEN Biotechnology Co. Ltd., Shanghai, China) was subsequently adopted. The HTR-8/SVneo cells were isolated, centrifuged, transfected for 45 h, and resuspended with the binding buffer. The cell suspension was stained with Annexin V-FITC/PI. The apoptosis rate was determined using the Attune™ NxT flow cytometer (Invitrogen, Thermo Fisher Scientific).
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