Figure 2.

Nuclear run-on analysis of APE-1 transcription. Untreated cells (lanes A) and cells treated with 130 nM HOCl (lanes B) for various times were harvested and their nuclei were isolated by a mild detergent lysis procedure (37, 38). Aliquots of nuclei were incubated for 30 min at 30°C with a transcription mixture containing [α-³²P]UTP, and the RNA was extracted by using RNAzol (Tel-Test, Friendswood, TX). The extracted RNA was hybridized to a nitrocellulose membrane on which 1 μg of APE-1 cDNA (Left) or β-actin cDNA (Right) was loaded in individual slots.