Journal: Nucleic Acids Research
Article Title: TDP1 repairs nuclear and mitochondrial DNA damage induced by chain-terminating anticancer and antiviral nucleoside analogs
Figure Lengend Snippet: Tdp1 −/− DT40 cells treated with CTNAs show enhanced perturbations in cell cycle profiles and greater DNA damage. ( A ) Wild-type and Tdp1 − / − DT40 cells were treated with 5 μM ACV or 150 nM Ara-C for 24 h before cell cycle analysis. ( B ) Western blots of wild-type and Tdp1 − / − DT40 cells treated with 2.5 μM ACV or 200 nM Ara-C probed for γH2AX are shown. β′-actin served as loading control, and the length of continuous drug treatment is indicated above each lane.
Article Snippet: Measurement of mitochondrial DNA copy number by quantitative real-time polymerase chain reaction DT40 Tdp1 − / − and wild-type cells were treated with indicated AZT or ddC (Sigma-Aldrich, St. Louis, MO, USA) concentrations, supplemented with 50 μM of uridine and incubated at 37°C for 24 h. The genomic DNA was extracted with Qiagen Dneasy Blood and Tissue Kit (Qiagen, Valencia, CA, USA) and used as templates in quantitative real-time polymerase chain reaction (PCR) with Power SYBR Green PCR Master Mix (Life Technologies, Carlsbad, CA, USA) in triplicates or quintuplets on an ABI 7900 thermocycler (Life Technologies, Carlsbad, CA, USA).
Techniques: Acetylene Reduction Assay, Cell Cycle Assay, Western Blot