Modfit lt software v5

HeLa cells were double blocked with 2 mM thymidine (arrested for 16 h, released for 8 h, and arrested for another 16 h), and then the cells were harvested for cell cycle and NEIL3 expression analysis at indicated time point. Cells were collected by digestion using trypsin and were washed twice with 1× cold-PBS. Cells were then fixed in 70% ethanol at −20°C overnight and stained with propidium iodide for 20 min. Cell cycle distribution was analyzed using a FACS Canto machine. The data was analyzed by FlowJo software v10.07 (BD Biosciences) or Modfit LT software v5.0 (Verity Software House).

After 48 h of transfection with control (Ctrl) or CEP295 siRNAs, U2OS cells were collected after digestion with trypsin and were washed twice with 1 × cold-PBS. The cells were then fixed in 70% ethanol at −20 °C overnight and stained with FxCycle™ PI/Rnase staining solution (Thermo Fisher Scientific #F10797) for 20 min. Cell cycle distribution was analysed using a FACS Canto™ flow cytometry system (BD Biosciences). The data were analysed using Modfit LT software v5.0 (Verity Software House).