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. Author manuscript; available in PMC: 2017 Mar 17.
Published in final edited form as: Mol Cell. 2016 Mar 17;61(6):850–858. doi: 10.1016/j.molcel.2016.02.010

Figure 1. End Joining in Xenopus Egg Extract Depends on Classical NHEJ Factors.

Figure 1

(A) Inhibition of end joining by immunodepletion of Ku70/80 with αKu80 antibody, and rescue with recombinant X. laevis Ku70/80. lin, linear DNA substrate; scc, supercoiled closed-circular products; oc, open-circular products. In other conditions, Ku immunodepletion selectively inhibited circularization, as previously reported (Fig. S1C) (Labhart, 1999; Di Virgilio and Gautier, 2005).

(B) Inhibition of end joining by the DNA-PK inhibitor NU-7441.

(C) DNA-PKcs immunodepletion.

(D) Immunodepletion of XLF (ΔXLF) or XRCC4 (ΔX4) and rescue with recombinant X. laevis XLF, XRCC4 (X4), wild-type LIG4:XRCC4 (L4X4), or catalytically inactive LIG4K278R:XRCC4 (L4X4 K278R). Lower panels in (A), (B), and (D) are western blots of immunodepleted extract with indicated antibodies. Uncropped blots are shown in Fig. S1F-H. XLF was not clearly visible in western blots of extract, but immunoprecipitated XLF was detected by western blotting and mass spectrometry (Fig. S1J-K).

See also Figure S1.