The cells were seeded at 5000 cells per well in 96-well plates and incubated overnight. After a particular treatment, the cell viability was determined using Cell Counting Kit-8 (Dojindo, Japan), according to the manufacturer’s instructions. The cell survival rates are expressed as the means ± SD from three independent experiments.
Apoptosis was examined by flow cytometric analysis. The cells were treated with certain concentrations of apatinib for the indicated durations. Both floating and adherent cells were collected, stained with Annexin V–fluorescein isothiocyanate (FITC), and propidium iodide (Dojindo, Kumamoto, Japan), and further analyzed with a flow cytometer (FACScan, BD Biosciences, San Jose, CA, USA) equipped with Cell Quest software (BD Biosciences). Apoptosis was also determined using the TUNEL (Terminal deoxynucleotidyl transferase dUTP nick end labeling) apoptotic cell detection kit (Roche, Basel, Switzerland), according to the manufacturer’s instructions. Apoptosis was expressed as the mean ± SD from three independent experiments.
Apoptosis was examined by flow cytometric analysis. The cells were treated with certain concentrations of apatinib for the indicated durations. Both floating and adherent cells were collected, stained with Annexin V–fluorescein isothiocyanate (FITC), and propidium iodide (Dojindo, Kumamoto, Japan), and further analyzed with a flow cytometer (FACScan, BD Biosciences, San Jose, CA, USA) equipped with Cell Quest software (BD Biosciences). Apoptosis was also determined using the TUNEL (Terminal deoxynucleotidyl transferase dUTP nick end labeling) apoptotic cell detection kit (Roche, Basel, Switzerland), according to the manufacturer’s instructions. Apoptosis was expressed as the mean ± SD from three independent experiments.