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. 2016 Apr 29;44(12):5743–5757. doi: 10.1093/nar/gkw326

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© The Author(s) 2016. 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/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — RPA suppresses MMEJ. (A) MMEJ repair efficacy of EJ2-GFP reporter cells treated with NT control siRNA (NT), POLQ siRNA (siPOLQ), RPA siRNA (siRPA) and POLQ plus RPA (siPOLQ+siRPA) indicated by the percentage of GFP-positive cells. Error bars show SEM from three independent experiments. **P < 0.01, ***P < 0.001, ****P < 0.0001. (B) POLQ gene expression measured by RT-qPCR using the primers indicated in Materials and Methods. (C) Western blot showing RPA knockdown 48 h following siRNA transfection. (D) Representative sequence alignments of the PCR products (see Supplementary Figure S13) in NT control (NT), and POLQ-depleted (siPOLQ) cells. I-SceI recognition sequence and terminal microhomologies at the break sites are highlighted. (E) Frequency of MMEJ at mis-repaired junctions in HPRT deletion mutants isolated from cells treated with NT control siRNA (NT) or POLQ siRNA (siPOLQ). P values calculated by statistical analysis ‘‘difference between proportions’’, * P < 0.05.