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Figure 3. Effect of xDNA2 on DNA end processing. (A) Depletion efficiency, end processing substrate, and reaction set up. The quantitation standards for western analysis were NPE loaded at the indicated amounts relative the depleted NPE. The 3′-ends of the partially filled-in linear pUC19 were labeled with 32P-dA by Klenow. (B) Effect of xDNA2 depletion on end processing. DNA (2 ng/µl) was incubated in xDNA2-depleted NPE, mock-depleted NPE or xDNA2-depleted NPE supplemented with either ELB buffer or the purified xDNA2 protein (to ∼15% of the endogenous xDNA2 level) in the presence of ddNTPs (to block NHEJ). Two additional reactions containing xDNA2 or ELB buffer served as controls. Samples were taken at the indicated time, treated with SDS/proteinase K and separated on a 1% TAE/agarose gel. The gel was first stained with SYBR Gold to detect total DNA and then dried for exposure to X-ray film to detect 32P. Image published in: Liao S et al. (2008) © 2008 The Author(s). This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license Larger Image Printer Friendly View

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