Modfit 4.0.5 lt cell cycle analysis software

Cells were seeded in six-well plates for three days in serum free medium (SFM). Cells were then placed in complete media (CM) before drug treatment for 24 h. After treatment, cells were trypsinized and suspended in 70% ethanol overnight. Cell suspensions were then pelleted, washed in PBS, and stained with a propidium iodide (PI) solution. Samples were then run using a NovoCyte flow cytometer (ACEA Biosciences, Inc., San Diego, CA, USA) and analyzed using NovoExpress 1.0.2 software. Samples were initially screened with SSC-H versus FSC-H density plot in linear scale and the gates were set to exclude debris. Another density plot was generated with the cells in the set gates and analyzed in PE-H versus PE-A for doublet discrimination. Singlet cells were gated and exported as FCS files and analyzed using ModFit 4.0.5 LT cell cycle analysis software (Verity Software House, Topsham, ME, USA).