Journal: PLoS ONE
Article Title: Mycoplasma fermentans Inhibits the Activity of Cellular DNA Topoisomerase I by Activation of PARP1 and Alters the Efficacy of Its Anti-Cancer Inhibitor
Figure Lengend Snippet: M. fermentans diminished the CPT inhibition effect on the DNA relaxation activity of Topo I. MCF7 cells were infected with M. fermentans ( M.f ) for 6 hrs followed by CPT (30 µM) treatments for additional 1.5 hrs. Total nuclear protein (12.5 ng) was added to a specific reaction mixture for Topo I. Reaction products were analyzed by agarose gel electrophoresis. ( A ) A representative picture n = 5, of Topo I DNA-relaxation activity. ( B ) Quantification analysis of Topo I activity. ( C ) Topo I protein level examined by Western blot analysis. Symbols: R and S are the relaxed and supercoiled form of the pUC19 DNA, respectively, Topo- no protein added to the reaction mixture t -test: * p
Article Snippet: Supercoiled DNA plasmid pUC19 was purchased from MBI Fermentas (Hanover, MD, USA).
Techniques: Cycling Probe Technology, Inhibition, Activity Assay, Infection, Agarose Gel Electrophoresis, Western Blot