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. Author manuscript; available in PMC: 2022 Oct 21.
Published in final edited form as: DNA Repair (Amst). 2021 Sep 1;108:103217. doi: 10.1016/j.dnarep.2021.103217

Fig. 6. RNF8 and RNF168 promote resection in Lig4−/−:53bp1−/− cells.

Fig. 6.

(A) Schematic of tetracycline-inducible CRISPR/Cas9-mediated gene inactivation in bulk abl pre-B cell populations using guide RNAs (gRNAs). Histogram of the flow cytometric analysis of FLAG-tagged Cas9 using FLAG antibody on permeabilized abl pre-B cells before (dashed line) and after (solid line) doxycycline (Dox) induction is also shown. (B) Western blot analyses with 53BP1 or RNF168 antibodies on cell lysate from Lig4−/− or Lig4−/−:53bp1−/− abl pre-B cells with chromosomally integrated tetracycline-inducible Cas9, expressing gRNAs targeting 53bp1 (g53bp1) or Rnf168 (gRnf168). (C) Southern blot analysis as described in Fig. 1C of XbaI-digested genomic DNA from imatinib-treated Lig4−/− or Lig4−/−:53bp1−/− abl pre-B cells with chromosomally integrated tetracycline-inducible Cas9, expressing guide RNAs targeting 53bp1 (g53bp1), Rnf8 (gRnf8) or Rnf168 (gRnf168) after 7 days of doxycycline treatment for gene inactivation. Percentages of intact SEs (iSEs) in imatinib-treated samples are listed below each lane.