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. 2017 Nov 16;46(1):229–241. doi: 10.1093/nar/gkx1103

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© The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 7. — RPA may be dispensable for synthesis by pol ϵ in the presence of E1. (A) Primed M13 template assays were performed with increasing amounts of RPA (3.5, 17.5, 35 and 70 ng/μl) in the presence of pol ϵ (lanes 11–14), pol ϵ and E1 (lanes 6–9) and pol ϵ and RFC/PCNA (lanes 17–20). Pol ϵ and E1 with no RPA serves as the positive control standard (lane 5). E1 alone (lane 3), pol ϵ alone (lane 4), RPA alone (lane 15) and RFC/PCNA alone (lane 21) were incubated with template and serve as controls, in which little (pol ϵ, lane 4) or no synthesis was observed. Lanes 1 and 2 represent reference size markers, as described in Materials and Methods. (B) Synthesis was quantified as in the Materials and Methods and graphed for three experiments. Results were quantified as in the Methods. Values for pol ϵ + E1 (dashed line) were plotted with pol ϵ + E1 with 0 ng/μl of RPA set to 100%. Values for pol ϵ + RFC/PCNA (solid line) were plotted setting pol ϵ + RFC/PCNA with 35 ng/μl of RPA to 100%, as this is the RPA level used for 100% synthesis of pol ϵ + RFC/PCNA throughout this manuscript and consistent with other published studies (11,20,21). Error bars are defined as s.d.