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. 2022 Oct 20;50(19):11058–11071. doi: 10.1093/nar/gkac913

Figure 3.

Figure 3.

Catalytically inactive Lig4 promotes joining of DSBs in episomal substrates in a variety of cell types. (A) Lig4−/− HCT116 cells were tested for TelN and I-SceI joining by co-transfecting the TelN/I-SceI substrates and expression plasmids with or without co-transfection of expression plasmids encoding either WT or catalytically inactive human ligase IV as indicated. (right) Immunoblot of cells transfected with indicated expression plasmids, and probed for either Lig4 or DNA-PKcs. (B) Lig4−/− U20S cells were tested for TelN and I-SceI joining by co-transfecting the TelN/I-SceI substrates and expression plasmids with or without co-transfection of expression plasmids encoding either WT or catalytically inactive mouse Lig4 as indicated. (right) Immunoblot of cells transfected with indicated expression plasmids, and probed for either Lig4 or DNA-PKcs. In A and B, student's T test comparing joining rates between vector and Lig4 or K273A were performed; ****P < 0.0001; ***P < 0.001; **P < 0.01; ns = not significant in two-tailed unpaired t test.