Research Paper Volume 11, Issue 9 pp 2852—2873

Poly(ADP-ribosyl)ation and DNA repair synthesis in the extracts of naked mole rat, mouse, and human cells

class="figure-viewer-img"

Figure 2. DNA synthesis and effect of PAR synthesis in WCEs. (A) DNA synthesis in the absence of NAD+. The cell extract proteins (0.5 mg/mL) were incubated for 5 min with 100 nM DNA duplexes bearing dRP, pDEG, or flap in the presence of 0.1 mM dNTPs (as described in the section ‘DNA synthesis assay’). (B) DNA synthesis in the presence of NAD+. The same as in (A), but in the presence of 0.5 mM NAD+. The unknown products are marked. Lanes 1 in A and B correspond to the initial primer (control). The types of DNA and cell lines are indicated. (C and D) Quantification of the products shown in Figure 2A and 2B, respectively. The white parts of the bars correspond to the non‐elongated primer, the grey parts reflect the amount of the primer elongated by one dNMP, and the black parts correspond to the products of strand‐displacement DNA synthesis. The intensity of the products is calculated as a percentage of the total radioactivity in the lane. The structures of DNA substrates are schematically shown at the top.