Annexin 5 fitc detection kit

The annexin V–FITC staining was performed as previously described [41 (link)]. Briefly, cells were incubated with indicated concentration of ILL for 24 h. The apoptotic cell death was examined using annexin V–FITC detection kits according to the manufacturer’s instructions (BD Biosciences, San Diego, CA, USA). The data (ten thousand events for each sample) were collected and analyzed by the BD Accuri C6 flow cytometer.

The cell cycle was determined by flow cytometry analysis as previously described [54 (link)]. Briefly, approximately 3 × 10⁵ cells were incubated with or without BEZ235 in the presence or absence of regorafenib for 48 h. Cells were stained with 20 μg/mL propidium iodide solution for cell cycle analysis, and apoptotic cell death was examined using annexin V-FITC detection kits according to the manufacturer’s instructions (BD Biosciences, San Diego, CA, USA). The data were collected and analyzed by the BD Accuri C6 flow cytometer.

Apoptosis of LO2 and HepG2 cells induced by PIX was detected by using Annexin V-FITC detection kits (BD Biosciences). Briefly, cells treated with PIX (16 to 32 μg/mL) for 24 h under standard conditions (DMEM with 10% FBS, 5% CO2, 37°C) were collected by centrifugation and resuspended gently in 100 μL of 1× Annexin V binding buffer at a concentration of 1 × 10⁶ cells/mL. Annexin V-FITC (5 μL) and PI (5 μL) were added to the suspension. After incubating at room temperature for 15 min in the dark, the stained cells were analyzed by flow cytometry (71 (link)).
For CLSM observation, RAW264.7 cells were cultured in standard conditions (DMEM supplemented with 10% FBS, 5% CO2, 37°C) supplemented with PIX (32 to 128 μg/mL) in humidified incubators. After incubating for 24 h, the cells were collected and stained with Annexin V-FITC detection kits. Then, 20 μL of the suspension was moved onto a glass slide, allowing it to be air dried and imaged.