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. 2000 Jul 11;97(15):8397–8402. doi: 10.1073/pnas.140137297

Figure 2.

Figure 2

Removal of 8-OxoG in MEF cell lines. Experimental scheme: Plasmid pSΔoriSV-[8-OxoG.C] has a deletion of the SV40 origin of replication (hatched) whereas pS(ori-p)SV-[8-OxoG.C] has a deletion of both the SV40 origin of replication and the SV40 promoter from −10 to 273 that eliminates transcription of TAg and of the 8-OxoG (hatched). After transfection in MEF cell lines, plasmid DNA was recovered and incubated with 5 ng of Fpg protein or left untreated before ethidium bromide-agarose gel electrophoresis and Southern blot analysis. (A) In this assay, removal of 8-OxoG is identified by the presence of CC plasmid DNA after incubation with Fpg (R; repaired). Unrepaired 8-OxoG causes conversion of the (CC) plasmid to an open circle (OC) when treated with Fpg (NR). (B) Control lanes (C) represent plasmid DNA (pSΔoriSV-[8-OxoG.C]) with (+) or without (−) Fpg treatment. Plasmid DNA transfected into wild-type or ogg1−/− MEF cell lines was incubated 2–12 h, as indicated and analyzed without Fpg treatment. The same result was obtained with pS(ori-p)SV-[8-OxoG.C].