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. Author manuscript; available in PMC: 2009 Mar 18.
Published in final edited form as: Methods. 2007 Feb;41(2):222–231. doi: 10.1016/j.ymeth.2006.07.024

Figure 2.

Figure 2

Aphidicolin induces hyperunwinding of plasmid DNA and Chk1 phosphorylation. Plasmid DNA was incubated in HSE for 30 minutes, and then transferred to an equal volume of NPE in the presence or absence of aphidicolin (final concentration 15 μM). 1 μL of each sample was analyzed for phospho-Chk1 (S344) or total Chk1 by immunoblotting at the indicated times (bottom panels). Replication was analyzed in parallel by incorporation of [α32P]-dCTP into plasmid DNA. DNA samples were analyzed via chloroquine agarose gel electrophoresis followed by SybrGold staining (top panel) and autoradiography (middle panel). [Reprinted with permission from Genes and Development, copyright April 15th, 2005.]