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. Author manuscript; available in PMC: 2015 Oct 1.
Published in final edited form as: Biochem Pharmacol. 2014 Aug 15;91(3):312–322. doi: 10.1016/j.bcp.2014.08.006

Fig. 5.

Fig. 5

Relationship between inhibition of DNA synthesis and accumulation of γH2AX. Panels A and B, HL-60 cells (4 × 105 cells/ml) were treated with various agents for 16 h (A) or for the times specified (B). Histones were extracted from 3 × 106 cells and subjected to western analyses using an anti-phospho-histone H2AX antibody. For a loading control, duplicate gels were stained with 0.05% Coomassie Brilliant Blue (A) or GelCode Blue Safe Protein Stain (Thermo Scientific, Lockford, IL) (B). Panel C, HL-60 cells (2 × 106 cells/ml) were treated with various agents for 1 h and labeled with 3H-thymidine for an additional 1 h. Radioactivity in the acid-insoluble fraction was determined. Panel D, HL-60 cells (4 × 105 cells/ml) were pretreated with 0.4 µM of triapine or 0.2 mM of HU for 1 h, washed and resuspended in fresh medium. At 0, 4, 8, and 16 h, cells were condensed to 2 × 106 cells/ml and labeled with 3H-thymidine for 1 h, and radioactivity in the acid-insoluble fraction was determined.