To assess cell proliferation, flow cytometric analysis was performed using PE-conjugated monoclonal anti-Ki-67 antibody (clone: 20Raj1, Bioscience) and apoptosis performed by Caspase 3 kit (BD Biosciences, San Diego, CA). For intracellular staining, cells were collected and washed twice with ice-cold PBS, resuspended with BD cytofix/cytoperm solution and incubated at 4°C for 20 minutes, cells were washed twice using Perm
Also, flow cytometric analysis was performed using Annexin V-FITC/PI staining. Following 48h treatment, cells were collected and rinsed twice with ice-cold PBS. Then cells washed and incubated with Annexin V-FITC/PI (eBiosciences) for 15 min at room temperature. Cells were analyzed by BD FACS Calibur with FlowJo (7.6.1) software for acquisition and analysis.