4.10. Immunofluorescence staining of DNA damage markers

Immunofluorescence staining was carried out as described previously [31]. Briefly, cells grown on coverslips were fixed in 4% paraformaldehyde for 10 min. The cells were then permeabilized in 0.2% Triton X-100 for 10 min, and blocked in 10% normal goat serum overnight at 4 °C. The coverslips were incubated with anti-phospho-H2AX,8-OHdG antibody overnight at 4 ℃,washed in PBS, and incubated with TRITC-conjugated Goat anti-mouse secondary antibody (Jackson Immuno Research Laboratories, West Grove, PA) for 1 h at room temperature. Cells were washed in PBS three times and counterstained with DAPI. Fluorescence images were captured under a fluorescence microscope.