Cellular DNA Content Analysis
The CyQuant-NF assay (C35006, ThermoFisher Scientific, USA), which measures cellular DNA content via fluorescent dye binding, was used to quantify the cell number in cultures treated with increasing doses of IFNγ. Briefly, control shRNA-expressing KCs or IFITM1 shRNA-expressing KCs were seeded 500 cell/well in 96-well plates and allowed to attach overnight. Cells were cultured in presence of indicated concentrations of IFNγ, in triplicate wells, for 96 h and then processed according to the protocol for adherent cells provided by the manufacturer. The fluorescence intensity detected is a measure for the number of cells present in the wells (34).