DT40 cells were grown in RPMI Medium 1640 (Life Technologies, 61870), supplemented with 7% fetal bovine serum (FBS, Life Techonologies, 10270), 3% chicken serum (Life Technologies, 16110), 50 µM β-mercaptoethanol and penicillin/streptomycin, at 37 °C in a 5% CO2 incubator. Sensitivity assays were performed as previously described6. In brief, 105 cells were incubated with drug-containing medium in a sealed FACS tube at 37 °C for 2 h (acetaldehyde) or 1 h (mitomycin C or cisplatin). Dilutions were plated in 6-well plates containing semi-solid medium (4000 cP methyl cellulose (M0512 Sigma), DMEM/F-12 powder (Life Technologies, 32500-043), 7% FBS, 3% chicken serum, 50 µM β-mercaptoethanol and penicillin/streptomycin). Plates were incubated for 7–10 days, after which timed colonies were counted manually. Survival is plotted as a percentage relative to untreated cells. Each data point represents the mean of three independent experiments each carried out in quadruplicate.
