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. Author manuscript; available in PMC: 2013 Nov 1.
Published in final edited form as: Free Radic Biol Med. 2012 Jul 27;53(9):1782–1790. doi: 10.1016/j.freeradbiomed.2012.07.025

Figure 5. Abasic site incision and nick ligation activities in nuclear extracts from DIV7 neurons are differentially affected by exposures to H2O2 and oligomycin.

Figure 5

Reactions were assembled with radioactively labeled substrates. Products resolved in denaturing polyacrylamide gels were visualized by autoradiography; representative autoradiograms are shown. Time points are indicated and a substrate-only lane is included (lane 1). (A) Incision assay of abasic site (THF) carrying substrate. Reaction with recombinant human Ape1 serves as positive control (lane 14). 5′-end labeled 20-mer substrate (S) and the 9-mer incision product (P) are indicated. (B) Nick ligation assay. 5′-end labeled 21-mer substrate (S) and ligated 45-mer product (P) are indicated. Labeled oligonucleotides resolved in parallel provide size markers (M). Bar graphs represent product yields calculated from phosphorimager values and expressed as pmoles/min/mg protein. Values for three or four independent assays for each substrate in the linear range of reaction were used to obtain mean ± SEM. *P<0.05 versus control.