Figure 2. CCAR2 depletion leads to hyper-recombination.

(a) Cells harbouring a single copy of SSR were transfected with the indicated siRNAs. A non-target siRNA (siNT) and an siRNA against CtIP were used as negative and positive controls, respectively. The resultant NHEJ:HR ratio was normalized to control cells and plotted. The average and s.d. of four independent experiments performed in triplicate are shown. Statistical significance is marked with one to three asterisks, as described in the Methods section. (b) Same as a, but with cells infected with lentiviral particles harbouring the indicated shRNAs. shNT (a non-target shRNA) and shRNA against CtIP were used as negative and positive controls, respectively. (c) Cell cycle distribution of cells after downregulation of the indicated genes with shRNA. (d) A schematic representation of the HR reporter DR-GFP is shown on the left side. An intramolecular gene conversion is shown, although it is possible that such reporter engages in an intermolecular recombination event with the sister chromatid, with the same results. Cells bearing such reporters were transformed with the indicated siRNAs. Further details are as in a. (e) Same as d, except that cells were transduced with viral particles containing the indicated shRNAs. (f) A schematic representation of the NHEJ reporter EJ5-GFP is shown on the left side. Cells bearing such a reporter were transformed with the indicated siRNAs. Further details are as in a.