A. 21-mer containing an 8-oxoG lesion was labeled at its 5' end using [32P] ATP and incubated with cortex and medulla kidney homogenate of wild type and Eker rat. Oligonucleotide cleavage products were analyzed on DNA sequencing gels and subjected to autoradiography. Pure human OGG1 enzyme (E) and buffer alone (S) were analyzed as positive and negative controls, respectively. The top arrow indicates the 21-mer of 8-oxodG as a substrate and the bottom arrow is the DNA cleavage product (13-mer). B. Histograms represent means ± SE (n = 3). C. DNA was extracted and digested with nuclease P1. The detection of dG and 8-oxodG was performed by HPLC-EC analysis. Authentic standards of 8-oxodG and dG were analyzed simultaneously. Standard curves for dG and 8-oxodG were prepared and quantitated by linear regression analyses. Significant difference from wild type rat is indicated by * < 0.05 and ** < 0.01.
Taken from "Tuberin haploinsufficiency is associated with the loss of OGG1 in rat kidney tumors"
Molecular Cancer 2008;7():10-10.
Published online 24 Jan 2008