MTT Assay

UWB1.289 MTT assays follow similar cell culture and transfection procedures as the comet assay until day five. On day five, or 48 hours after the second transfection, 2000 cells of each treatment were counted and plated in a 96-well plate (~0.143 cm²). Cells were cultured for 72 hours in a 96 well plate.

Primary ovarian cancer cells were counted and 1000 cells were plated per well of a 96-well tissue culture plate. 0, 5, 10, and 20 µM cisplatin and 0 and 75 nM TSA were applied to both platinum sensitive and platinum resistant primary ovarian cells.

Three days after the start of the MTT assay, 10 µl of 12 mM MTT stock solution (~10% of the well volume) was added to each well. The cells were incubated at 37 °C for four hours, after which cell membranes were disrupted with a 10% sodium dodecyl sulfate (SDS), 0.01 M hydrochloric acid (HCl) solution. After a final four-hour incubation at 37 °C, the plate is shaken to ensure mixing and the absorbance is measured at 600 nm. Each treatment was performed in triplicate.