(A) Cut linear DNA, supercoiled circular DNA, or buffer was added to extract containing S-labeled Ku80

After incubation at 22°C for the indicated times, samples were taken. The asterisk indicates DSB-dependent modified Ku80. Radioactively labeled protein was visualized using a phosphorimager. (B) Quantification of the radioactivity of the entire lane in A using a phosphorimager ( = 4). (C) Degradation, quantified as in B, of Ku80, Ku70, and Mre11 in the presence of cut DNA ( = 3). (D) Degradation, quantified as in B, of Ku80 in the presence of cut DNA and buffer, 0.5 mg/ml wild-type ubiquitin, or 0.5 mg/ml ubiquitin-K48R ( = 3). (E) Degradation of Ku80 in the presence of buffer, supercoiled DNA, or cut DNA. At indicated time points extract samples were spotted onto glass microfiber filters and precipitated in 10% TCA, and radioactivity was quantified using a liquid scintillation counter ( = 4). (F) Degradation, quantified as in E, of Ku80 in the presence of either DMSO or 100 μM MG132 ( = 4). Error bars denote one standard error of the mean.Copyright information:Taken from "Ku80 removal from DNA through double strand break–induced ubiquitylation"The Journal of Cell Biology 2008;182(3):467-479.Published online 11 Aug 2008PMCID:PMC2500133.