1. Ligation efficiency is abrogated with increasing concentration of antibodies against NHEJ proteins.
Ligation efficiency is abrogated with increasing concentration of antibodies against NHEJ proteins. Agarose gels of ligated pUC18 after incubation with nuclear extracts containing antibodies (1:200–1:10; Lanes 2–6) to Ku86 or Ku70. Results for the HL60 cell line (A), an AML sample (C), and a normal CD34+ sample (E). A portion (3 μg) of nuclear extract per sample was used (M, monomer; D, dimer; T, trimer; Te, tetramer). B, D, and F, ligation efficiency with increasing concentration of antibodies to NHEJ proteins incubated with nuclear extracts from HL60 cells (B), AML cells (D), and normal CD34+ cells (F). Anti-Ku86 ( ), anti-DNA-PKcs (▴), anti-Ku86 + blocking peptide (····), anti-Ku70 (▪) and anti-Rad51 control (•). G, agarose gel of ligated pUC18 after incubation of nuclear extracts containing a 1:10 dilution of anti-Ku86 antibody. Lane 1, pUC18 ligation in a CML sample with no anti-Ku86 added; Lanes 2–4, CML samples (n = 3); Lanes 5–7, AML samples (n = 3); Lane 8, K562; Lanes 9–10, normal PBLC samples (n = 2); Lanes 11–12, normal CD34+ samples (n = 2). A portion (3 μg) of nuclear extract per sample was used.
Terry J. Gaymes et al. Cancer Res 2002;62:
©2002 by American Association for Cancer Research