The formation of γ-H2AX and 8-oxodG after exposure of fibroblast Balb/c-3T3 cells with N-OHAAF/UV irradiation as measured by double immunofluorescence staining. (a) Representative images of Balb/c-3T3 cells stained for Hoechst 33342 (nuclear stain), monoclonal antibodies of γ-H2AX and 8-oxodG under different conditions as indicated. (b and c) Quantitative results of γ-H2AX and 8-oxodG formation in Balb/c-3T3 cells under different conditions as indicated. **Significant difference from the control groups, P < 0.001. The intensity and amount of γ-H2AX and 8-oxodG immunoreactivity were quantified by CLSM quantitative software. All groups were first pretreated with the indicated chemicals in serum-free DMEM medium (high glucose, no glutamine, no phenol red) for 1 h and then under dark or UV irradiation (313 nm, 1.5 mW/cm2) treatment for 20 min. The solvent control group was conducted with 0.15% acetonitrile. The 20 mM N-OHAAF mother liquor was prepared by dissolving the synthesized solid in acetonitrile.
