Celigo® cytometer imaging assay

FaDu and CNE-2Z cells infected with lentiviral vectors (shNUDT21 or shCtrl) were seeded in 96-well plates at a density of 2×10⁵ cells per well. Two types of green fluorescent cells were detected using a Celigo® cytometer (Nexcelom, Inc., USA). We accurately counted the number of green fluorescent cells per scan well by adjusting the assay setup input parameters. The ratio of the cell count at each time point of each group to the cell count at the first day of this group was calculated to obtain the cell proliferation multiple at each time point of this group, and growth curves based on cell proliferation folds were plotted according to cell proliferation folds and time points. We observed the control group and NUDT21 siRNA cell number for 5 days, calculated and analyzed the cell number, and constructed the cell proliferation curve.