Annexin 5 fitc 7 aminoactinomycin d 7aad

Cell viability was determined using annexin V-FITC/7-aminoactinomycin D (7AAD) staining from BD Biosciences according to the manufacturer’s instructions. SW620, SW480, and HT29 cells were seeded in 12-well plates and allowed to attach overnight. The following day, cells were treated with 2.5, 5, and 10 µM of Xn for 24 and 48 h. Floating cells and adherent cells were harvested, washed twice with cold PBS, and then incubated with 5 µL of annexin V-FITC and 5 µL of 7AAD in 50 µL of binding buffer for 15 min at room temperature in the dark. At the end of the incubation, 200 μL of binding buffer was added and cells were kept on ice until analysis. Compensation controls included unstained cells and cells stained with either annexin V-FITC or 7AAD alone.

Cell viability was determined using annexin V-FITC/7-amino-actinomycin D (7AAD) staining from BD Biosciences, as previously described [21 (link)]. Briefly, MES-SA and MES-SA/Dx5 cells were seeded in 12-well plates. The following day, cells were treated with 12.5, 25, and 50 µg/mL of PC and TR for 72 h. Floating and adherent cells were incubated with 5 µL of annexin V-FITC and 5 µL of 7AAD in 50 µL of binding buffer for 15 min at room temperature in the dark. Thereafter, 200 μL of binding buffer was added and cells were analyzed with a BD FACSCanto^TM cytometer (BD Biosciences, Le Pont de Claix, France).