Cell viability assays for Pitstop 2 treatment

A total of 2000 cells were seeded in 100 μl of medium in 96-well plates and incubated overnight at 37 °C. Two-fold serial dilutions of Pitstop 2 (Abcam) in medium were prepared with a DMSO control. An aliquot (50 μl) of drug or DMSO preparation was added to the cells and incubated at 37 °C for 4 days when cell viability was quantified using the CellTiter-Glo assay, as described. Survival fractions were calculated by dividing the raw luminescence values for the drug treated well by the DMSO controls. Data were analysed using GraphPad Prism and survival curves determined.