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. Author manuscript; available in PMC: 2018 Apr 17.
Published in final edited form as: Chem Res Toxicol. 2017 Mar 29;30(4):980–995. doi: 10.1021/acs.chemrestox.6b00389

Figure 1.

Figure 1

Concentration-dependent formation of DPCs in nuclear protein extracts prepared from HeLa human cervical carcinoma cells following exposure to cisplatin. (A) Nuclear protein extracts from HeLa cells (500 μg) and 5′-biotinylated double-stranded oligodeoxynucleotides (3.12 nmol) were incubated in the presence of 0–50 μM Cisplatin. The resulting DPCs were captured on streptavidin beads, and the proteins were resolved on 12% SDS-PAGE. Gels were stained with SilverQuest SilverStain to visualize the cross-linked proteins. (B) Densitometric analysis of protein bands in the 25 – 250 kDa molecular weight range was used to estimate the extent of total protein cross-linking to DNA in the presence of cisplatin. Known amounts of nuclear protein extract were analyzed as a control to estimate the cross-linking efficiency.