Assessing siRNA-Transfected Cell Viability

The viability of siRNA-transfected VK2/E6E7 cells was assessed using a 2,5-diphenyl-2H-tetrazolium bromide (MTT) assay [41 (link)]. Cells were seeded at a density of 1 × 10⁴ cells per well in 96-well culture plates and were allowed to adhere overnight. The following day, the culture medium was discarded and replaced with a medium containing 10% MTT (Sigma Aldrich). Cells were then incubated for 3 h at 37 °C. Afterward, the MTT solution was carefully removed from each well, and dimethyl sulfoxide (DMSO; Bio Basic Inc., Markham, ON, Canada) was added to dissolve the formazan crystals. The absorbance of each sample was subsequently measured at 570 nm using a microplate spectrophotometer (Multiskan Go; Thermo Fisher Scientific).