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. 2020 Jul 12;25(14):3180. doi: 10.3390/molecules25143180

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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(A) Western blot gel of the Topo II-DNA cleavage assay. Lane 1-2: control group of supercoiled pBR322 DNA without or with Topo II; Lane 3-5: addition of etoposide (100 μM), 4d or 4n (50 μM) on Topo II with supercoiled pBR322 DNA; Lane 6-7: pretreatment of pBR322 DNA and Topo II with 4d or 4n (50 μM) followed by addition of etoposide. In each lane, the positions of supercoiled DNA (S), relaxed DNA (R), linear DNA (L) and nicked DNA (N) are indicated. (B) Semi-quantification of the western blot gel in panel A. Data were presented as Mean ± SD (n = 3). * p < 0.05 indicates statistically significant difference of relaxed DNA compared with group E (Student’s t-test). (C) The unwinding assay of compounds. Lane D: pBR322 DNA; lane T: pBR322 DNA + Topo I; other lanes: pBR322 DNA + Topo I + 4d, 4n or EB of different concentrations.