FIGURE 11.

Detection of ADP-ribosylation in plant cells. (A) Western blot analysis of protein ADP-ribosylation in WT and mutant plant extracts. Wild-type Col-0 and AP endonuclease-deficient ARP^–/– mutant seeds were grown on MS agar medium. Two-week-old Arabidopsis plants were transferred to 10 μg•ml^–1 of bleomycin for 18 h. Total proteins were extracted from whole plants, separated by SDS-PAGE, and analyzed by immunoblotting using anti-PAR and anti-atPARP2 antibodies. Arrows indicate PARylated proteins and atPARP2 protein. (B) Detection of PAR–DNA adducts in gDNA extracted from 14-day-old seedlings grown under either normal conditions or genotoxic stress. Different quantities of gDNA in TE buffer (10 mM Tris-Cl pH 8.0, 1 mM EDTA) were spotted onto a nylon membrane, followed by mouse monoclonal anti-poly(ADP-ribose) antibody 10H dot blotting. For more details, see section “Materials and Methods.”