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. 2019 Apr 4;15(4):e1008101. doi: 10.1371/journal.pgen.1008101

Fig 3. Effect of LIG4 deficiency on Cas9-mediated switching and der3 using blunt, 5’, and 3’DSBs in human B cells.

Fig 3

(A) Percentage of IgA+ wild-type (WT) or LIG4-/- BJAB cells using blunt, (B) 5’DSBs, or (C) 3’DSBs as measured by flow cytometry, 5 days post-transfection with Cas9/sgRNA plasmids. (D) Frequency of Cas9-mediated der3 in WT or LIG4-/- BJAB cells using blunt, (E) 5’DSBs, or (F) 3’DSBs as measured by qPCR. Cells were harvested 5 days post-transfection with Cas9/sgRNA plasmids for genomic DNA isolation. (G) Total resection of S’μ and BCL6 at der3 junctions from 5’DSBs generated in WT and LIG4-/- BJAB cells. Dotted line denotes the total overhang length in S’μ plus BCL6 (95 nt). (H) Microhomology usage at S’μ-BCL6 junctions from 5’DSBs generated in WT and LIG4-/- BJAB cells. Black line denotes mean microhomology usage.