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. 2013 May 27;201(5):693–707. doi: 10.1083/jcb.201302145

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© 2013 Reczek et al.

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Figure 7. — Cells lacking the Brca1–Ctip interaction are proficient for DSB repair by HDR. ES cells harboring the DR-GFP substrate (A) integrated into the Pim1 locus were transfected with either an I-SceI expression vector or the empty vector. (B) I-SceI expression induced the appearance of GFP-positive cells to comparable levels in cultures of isogenic Ctip^+/− (subclones 1 and 2) and Ctip^S326A/− (subclones 3 and 4) ES cells, indicating that the Ctip-S326A mutation does not appreciably affect HDR. In contrast, a Brca1 mutant ES cell line 236.44 (Brca1^{Δ223–763/Δ223–763}) with the DR-GFP substrate integrated at the same position of the Pim1 locus (subclone 5) was deficient for HDR, as previously reported (Moynahan et al., 1999). Each ES subclone was assayed in triplicate with three independent transfections, and the error bars represent SEM. Similar results were also observed in separate experiments using additional independently derived Ctip^+/− and Ctip^S326A/− ES subclones.