Annexin 5 fluorescein isothiocyanate fitc kit

Cell apoptosis was determined by using the Annexin V-fluorescein isothiocyanate (FITC) kit (Thermo Fisher Scientific Inc.). After exposure to Pt(II) compounds at the IC₅₀ concentration for 24 h, YAPC cells were collected, washed twice with PBS, and subjected to centrifugation at 1200 × g for 5 min at room temperature. Subsequently, the cell pellet was resuspended in Annexin V-FITC and propidium iodide (PI) solutions. After incubation for 15 min at room temperature in the dark, additional Annexin V binding buffer (10 mM Hepes, 140 mM NaCl, and 2.5 mM CaCl₂) was added to each sample. The cells were analyzed using a flow cytometer (BD Biosciences, San Jose, CA, USA) and BD Accuri C6 Software.

The Annexin V-fluorescein isothiocyanate (FITC) kit (Thermo Fisher Scientific, Inc.) was used to determine cellular apoptosis. After exposure to curcumin, Cisplatin, or Pt-Curc NCs at the IC₅₀ concentration for 24 h in 37°C, MCF7 cells were collected, washed twice with PBS, and subjected to centrifugation at 1200 × g for 5 min at room temperature. Subsequently, the cell pellet was resuspended and treated with Annexin V-FITC and propidium iodide (PI) solutions. After incubating for 15 min at room temperature in the dark, additional Annexin V binding buffer (10 mM Hepes, 140 mM NaCl, 2.5 mM CaCl₂ in ddH₂O) was added to each tube, and the cells were analyzed.
To analyze the apoptosis in 3D spheroids, 5-day-old spheroids were chosen and exposed to curcumin, Cisplatin, or Pt-Curc complex NCs at the IC₅₀ concentration, for 24 h at 37°C. After the incubation period, all spheroids were rinsed with 10 mL of PBS and were collected in a 15 mL tube. After centrifugation (5 min, 400 g), spheroids were trypsinized to obtain a single cell suspension by gentle pipetting. Subsequently, the cell pellet was resuspended and treated with Annexin V-FITC and propidium iodide (PI) solutions.
Flow cytometry was performed on a flow cytometer (BD Biosciences, San Jose, CA, USA) equipped with FlowJo software.