2.6. Flow cytometry

The transfected Caki-1 and 786-O cells were harvested and trypsinized. Subsequently, the precipitate was washed in phosphate-buffered saline (PBS; Solarbio) and fixed with ethanol for 1 h. After incubation with RNase (Seebio) for 30 min, the cells were stained with propidium iodide (PI; Abcam, Cambridge, UK). Finally, cell distribution was monitored by FACScan Flow Cytometry (BD Biosciences, San Diego, CA, USA).

Cell apoptosis was assessed using Annexin V-FITC/PI Apoptosis Detection kit (Vazyme, Nanjing, China) following the manufacturer’s instructions. The apoptosis cells were measured by FACScan Flow Cytometry (BD Biosciences).