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. 2021 Sep 3;10:e68466. doi: 10.7554/eLife.68466

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Figure 3. — (A) Cas9-induced Lig4⁻^/−:Trp53bp1⁻^/− and Lig4⁻^/−:Lin37⁻^/− abl pre-B cells with (+) and without (−) the Ctip gRNA (gCtip). Western blot analysis with indicated antibodies (left) and flow cytometric analysis of chromatin-bound RPA before and after IR of the non-cycling cells (right) are shown. Representative of three experiments. (B) End-seq tracks of representative AsiSI sites on mouse chromosomes 1, 7, and 19 in non-cycling Lig4⁻^/− and Lig4⁻^/−:Lin37⁻^/− abl pre-B cells. (C) The heatmaps of End-seq at AsiSI DSBs across the mouse genome (y-axis) after AsiSI induction in non-cycling Lig4⁻^/− and Lig4⁻^/−:Lin37⁻^/− abl pre-B cells. Two experiments were carried out in two independently generated Lig4⁻^/−:iAsiSI and Lig4⁻^/−:Lin37⁻^/−:iAsiSI clones. DSB, double-strand break; End-seq, End Sequencing; gRNA, guide RNA; IR, ionizing radiation.

Figure 3—figure supplement 1. — (A) Cas9-induced Lig4⁻^/−:Trp53bp1⁻^/− and Lig4⁻^/−:Lin37⁻^/− abl pre-B cells with (+) and without (−) the Ctip gRNA (gCtip). Western blot analysis with indicated antibodies (left) and flow cytometric analysis of chromatin-bound RPA before and after IR of the non-cycling cells (right) are shown. Representative of three experiments. (B) End-seq tracks of representative AsiSI sites on mouse chromosomes 1, 7, and 19 in non-cycling Lig4⁻^/− and Lig4⁻^/−:Lin37⁻^/− abl pre-B cells. (C) The heatmaps of End-seq at AsiSI DSBs across the mouse genome (y-axis) after AsiSI induction in non-cycling Lig4⁻^/− and Lig4⁻^/−:Lin37⁻^/− abl pre-B cells. Two experiments were carried out in two independently generated Lig4⁻^/−:iAsiSI and Lig4⁻^/−:Lin37⁻^/−:iAsiSI clones. DSB, double-strand break; End-seq, End Sequencing; gRNA, guide RNA; IR, ionizing radiation.