Annexin 5 fluorescein isothiocyanate fitc propidium iodide detection kit

To determine myocardium apoptosis levels, cultured HL-1 cells were collected after transfection and rinsed in chilled PBS, followed by double staining with an Annexin V fluorescein isothiocyanate (FITC)/propidium iodide (PI) detection kit (Invitrogen) in the dark for 15 min. Then, the rates of apoptotic cells were detected by a flow cytometer (Beckman Coulter, Brea, CA, United States).

The extent of apoptosis was evaluated using an Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) detection kit (Invitrogen). 786-o and ACHN were cultured at 37°C and 5% CO₂ in six-well plates and transfected with miR-184 mimic or negative control at a confluence of approximately 60%. For apoptosis assays, floating and adherent cells were collected 48 h after transfection and then combined and washed twice with pre-chilled PBS and resuspended in in 1X binding buffer (Invitrogen). In total, 5 μl Alexa Fluor^® 488 Annexin V (Invitrogen) and 3 μl PI (Invitrogen) were added to 500 μl cell suspension, and the samples were analyzed within 15 min of staining. The fluorescence was analyzed by flow cytometry (Beckman Coulter, Miami, FL, USA) using an excitation of 488 nm, according to the manufacturer’s instructions. Each experiment was performed at least three times.