4.10. Cell Cycle Analysis

Cells were seeded at 70% confluence at 37 °C and 5% CO₂. After 24 h, cells were treated with THZ1, tamoxifen for 48 h or CDK7 siRNA-2 for 72 h. After the treatment incubation, the floating cells were aspirated and discarded. The cells were detached using trypsin then washed two times with cold PBS, and centrifuged. The pellet was resuspended with PI for 20 min for flow cytometry analysis. Flow cytometry was performed with a FACScan flow cytometer (Becton Dickinson). A minimum of 10,000 cells/sample were collected, and cell cycle analysis was conducted with the resulting DNA histograms using ModiFitLT software (Verily Software House, Topsham, ME, USA).