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. 2017 Sep 28;45(20):11782–11799. doi: 10.1093/nar/gkx871

Figure 2.

Figure 2.

3E10 scFv proteins that do not bind DNA still inhibit HDR. (A) Primary fibroblasts were transfected with the 2XMBP-scFv expression constructs. Cellular expression of each scFv was verified via immunofluorescence. The effect of each scFv on double strand break repair pathways was assayed using a luciferase plasmid-based host cell reactivation assay. (B) Schematic of the pre-digested luciferase constructs used to assay homology-directed repair (HDR). (C) Primary skin fibroblasts were transfected with 2XMBP-scFv expression constructs and then transfected with the pre-digested luciferase reporter constructs. (D) Results from the luciferase plasmid-based host cell reactivation assay, plotted as relative HDR (normalized to cells transfected with the luciferase constructs alone) after transfection with scFv expression constructs. (E) Primary skin fibroblasts were pre-treated with purified scFv proteins and then transfected with the pre-digested luciferase reporter constructs. (F) Results from the luciferase plasmid-based host cell reactivation assay, plotted as relative HDR (normalized to cells transfected with the luciferase constructs alone) after treatment with purified 2XMBP-scFv. Error bars represent the SEM; **P <0.01 and *P <0.05 by unpaired t-test.