Facs verse facsuite software

Cell cycle arrest after shRNA treatment was evaluated by PI staining as described earlier [16 ]. Briefly, after 48 hr of shRNA treatment cells were washed and fixed in 70% ethanol overnight. Next day cells were treated with PI stain (1 mg/ml) and RNAse solution and kept in dark at 37°C for 30 min before acquisition. Acquisition and analysis was done using BD-FACS CALIBUR (BD Biosciences, California, USA) and BD FACS Verse BD FACSuite software (BD Biosciences San Jose USA). Caspase mediated DNA nicking was analyzed using Apo-BrdU- Red in-situ DNA fragmentation assay kit (Biovision, Milpitas, CA, USA) as described earlier [16 ]. Acquisition and analysis was done using BD FACS Verse BD FACSuite software (BD Biosciences San Jose USA).