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. 2002 Apr 30;99(10):6860–6865. doi: 10.1073/pnas.092662499

Figure 3.

Figure 3

Overexpression of AAG in β-pol null cells. (a) Detection of AAG protein by immunoblotting of whole-cell extracts from 20 μg β-pol null(Neo) cells (Left) and 20 μg β-pol null(AAG) cells (Right). The arrow indicates the AAG band. (b) AAG activity in whole-cell extracts. The relative functional expression of AAG in the β-pol null(Neo) and β-pol null(AAG) cells was determined by release of hypoxanthine from a single lesion-containing 24-mer double-stranded oligonucleotide, as described in Materials and Methods. Lanes 1–4: β-pol null(Neo) cell extract incubated for 0, 2, 4, and 6 min, respectively. Lanes 5–8: β-pol null(AAG) cell extract incubated for 0, 2, 4, and 6 min, respectively. Lane 9: no extract control incubated for 6 min. (c) Methylation-induced increase in mutant frequency at λ cII. Spontaneous (filled bar) and MMS-induced (open bar) mutant frequencies at λ cII in β-pol null(Neo) cells or β-pol null(AAG) cells. Mutant frequencies were calculated after analysis of 3.2 × 106 pfu [β-pol null (Neo); spontaneous], 2.9 × 106 pfu [β-pol null (Neo); +1 mM MMS], 3.1 × 106 pfu [β-pol null (AAG); spontaneous], and 3.2 × 106 pfu [β-pol null (AAG); +1 mM MMS], respectively.