Fc500 flow cytometry system with cxp software

Cell cycle analysis was performed using a commercial kit (Coulter DNA Prep™ Reagents Kit; Beckman Coulter, Fullerton, CA, USA). Cells were plated in six-well plates and incubated overnight before treatment (Mock, 50 nmol/l dsCon and 50 nmol/l dsPAWR-435). Following treatment, cells were harvested, then washed twice with pre-chilled PBS and resuspended in 100 μl PBS at a concentration of 1×10⁶ cells/ml. Each cell sample was mixed with 100 μl DNA Prep LPR (contained in Coulter DNA Prep™ Reagents Kit), gently mixed by vortex and incubated in the dark at room temperature (25°C) for 20 minutes. Then each was mixed with 1 ml of stain (DNA Prep Stain; contained in Coulter DNA Prep™ Reagents Kit), gently mixed by vortex and again incubated in the dark at room temperature (25°C) for 20 minutes. Finally, cell cycle analysis was performed within 1 hour using flow cytometry (Beckman Coulter FC500 Flow Cytometry System with CXP Software; Beckman Coulter, Fullerton, CA, USA), and the raw data was analyzed by Multicycle for Windows (Beckman Coulter).

Cell cycle analysis was performed using the Coulter DNA Prep™ Reagents Kit. Cells were plated in six-well plates and incubated overnight before treatment. After treatment for 24h, harvested cells were washed twice with prechilled PBS and resuspended with 100 μl PBS at a concentration of 1×10⁶ cells/ml. Each cell sample was mixed with 100 μl DNA Prep LPR, gently mixed with a vortex, and incubated in the dark at room temperature (25°C) for 20 minutes. Then, each sample was mixed with 1 ml of DNA Prep Stain, mixed gently with a vortex, and incubated again in the dark at room temperature (25°C) for 20 minutes. Finally, the flow cytometry system (Beckman Coulter FC500 Flow Cytometry System with CXP Software; Beckman Coulter, Fullerton, CA, USA) was used for cell cycle analysis within 1 hour, and raw data was analyzed by Multicycle for Windows (Beckman Coulter).