2.10. Cell proliferation assay

Cells were seeded at a density of 1x10⁵cells/cm² and incubated for 24 and 48 h after transfection before cell proliferation was assayed. MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide) was used to measure cellular proliferation (Sigma Aldrich, Gillingham, UK)(Mosmann, 1983). MTT was dissolved in PBS at 5 mg/ml and filtered-sterilised and then added to cells at 24 h and 48 h. Cells were incubated at 37 °C for 3 h. After 3 h, the media was removed and 0.04 N HCL-isopropanol was added to solubilise the converted formazan. Absorbance of formazan was measured at a wavelength of 570 nm with a background absorbance of 630-690 nm. t-Tests were performed on the MTT data using R.