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. 2004 Jul 6;32(12):3531–3536. doi: 10.1093/nar/gkh676

Figure 4.

Figure 4

Immunoprecipitation of APE1 from gel filtration fractions containing 3′-PG activity. Fractions from gel filtration chromatography containing 3′-PG activity (fractions 19–22) were mock-immunodepleted and immunodepleted of APE1 using APE1-specific antibodies. Samples were analysed by SDS–PAGE and western blotting using antibodies against APE1 and Pol β (A). The original fractions, mock-immunodepleted and immunodepleted fractions 20 and 21 were tested for 3′-PG activity using a 5′ end labelled 3′-PG oligonucleotide annealed to an oligonucleotide (51mer) containing a hairpin loop and a 5′-phosphate residue. Samples were incubated for 10 min at 30°C in the absence of dNTPs prior to the addition of formamide loading dye and analysis by 20% denaturing PAGE and phosphorimaging (B).