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. Author manuscript; available in PMC: 2017 Oct 20.
Published in final edited form as: Cell Rep. 2017 Sep 19;20(12):2810–2819. doi: 10.1016/j.celrep.2017.08.091

Figure 3.

Figure 3

Effect of LIG4 insert1 on cellular joining of complex end structures (A) Cells were engineered to express LIG4ΔI from the native LIG4 locus by CRISPR/Cas9-based gene targeting, then the LIG4ΔI-a clone was reverted back to wild-type (LIG4+r/+r) by a second round of gene targeting (B–E) Substrates with varied end structures were introduced in the cell types described in (A). Joining efficiency was assessed by qPCR, and product structure by sequencing or diagnostic restriction digestion and defined as directly ligated or ligated after end processing as noted. Cellular NHEJ was assessed for (B) complementary ends, (C) ends with 3’ G:T terminal mismatches, (D) ends with 5’ terminal 8-oxoguanine (Go; product structures reported in Supplementary Figure 3D), and (E) ends with fully non-complementary overhangs (product structures reported in Supplementary Figure 3E). Error bars represent the standard error of the mean for 3 experiments. Means of linearized qPCR data and direct joining products were assessed by one-way ANOVA as significantly different from control (LIG4+/+ cells) with confidence p<0.001 (***) or not significantly different (ns).