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. 2017 Oct 20;45(20):11837–11857. doi: 10.1093/nar/gkx847

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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 8. — FANCD2 plays a crucial role to promote HDR-mediated, RAD51-dependent DNA DSB repair. (A) A GFP-HDR DNA repair assay was used to determine the HDR efficiency in WT, D2^−/−(clones #29 and #39), I^−/− (clones #28 and #30) and ID2 DKO (clones #1 and #2), as well as in the complemented cells. RAD54B^−/−/− cells were included in the assay as a control cell line that is severely HDR deficient. In this assay, I-SceI restriction enzyme digestion creates a DSB in the HDR reporter plasmid (DR-GFP). Repair of the DSB by HDR restores GFP expression. The repair efficiency was determined by dual GFP and mCherry positive cells divided by the mCherry positive cells (transfection control). Results were averaged from a minimum of three replicates and normalized to the average repair efficiency in the WT cells. Data points were averaged between clones of identical genetic backgrounds. Statistical significance at P < 0.05, P < 0.01, P < 0.001 and P < 0.0001 are indicated as *, **, ***, **** respectively. (B) The HDR assay shown in (A) was repeated for the WT, D2^−/−, I^−/− and ID2 DKO cells in the absence or presence of the RAD51 stabilizer, RS-1 (7.5 mM). Results were averaged from a minimum of three replicates and normalized to the average repair efficiency in the WT cells. Statistical significance at P < 0.05, P < 0.01, P < 0.001 and P < 0.0001 are indicated as *, **, ***, **** respectively.