Figure 4. An-Hq₂ Modifies dsDNA In Vitro.

(A) Primer extension from a 392-nucleotide dsDNA resulted in a full-length extension product. Controls lanes include: lacking oxidant (−OX), incubation with the end products (EP), a pre-quenched reaction (PQ), and a negative control containing DMSO (NR). Control lanes did not alter the amount of full-length product produced. Reaction with either cisplatin (CIS) or An-Hq₂ (RXN) in the presence of an oxidant led to a loss of full-length extension. (B) Gel showing sequence dependent damage. Sequence lanes on left. Unmodified DNA and controls gave similar patterns of extension stops. DNA incubated with cisplatin led to extension stops at polyguanine sequences. In contrast, An-Hq₂ and oxidant caused damage at most sequences. (C) Quantification of An-Hq₂ damage. Extension stops in the reaction (black) and in the pre-quenched control (light grey) were compared to a thymine sequence lane (T, dark grey). A decrease in extension stops was observed at thymine sequences.