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. 2012 Feb 27;7(2):e32313. doi: 10.1371/journal.pone.0032313

Figure 1. HAP treatment leads to the appearance of EndoV sensitive sites in HeLa DNA.

Figure 1

We extracted genomic DNA from HeLa cells grown with or without HAP. Treatment of this DNA with bacterial EndoV creates 3′ nicks, which are substrates for nick-translation (BioProbe® Nick translation kit with bio-16-dUTP (Enzo Life Sciences)) as described in Materials and Methods. A. Agarose gel electrophoresis of nick-translated DNA from HeLa cells. 1- from untreated cells; 1a – from untreated cells digested with DNase; 2 – from cells grown in 2.64 mM HAP; 3- from untreated cells, DNA incubated with Endo V; and 4 - from cells grown in 2.64 mM HAP, DNA incubated with Endo V. B. Detection of newly synthesized biotinylated DNA separated by alkaline agarose electrophoresis. 1- from untreated cells; 2 – from cells grown in 2.64 mM HAP; 3- from untreated cells, DNA incubated with Endo V; and 4 - from cells grown in 2.64 mM HAP, DNA incubated with Endo V.