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. 2010 Feb 15;207(2):417–427. doi: 10.1084/jem.20092449

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© 2010 Boboila et al.

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Figure 5. — The Ku-dependent A-EJ pathway uses a distinct mechanism for S region joining. (a) Classical NHEJ performs both direct (left) and MH-mediated (right) end-joining. Direct joining occurs when DNA ends are joined either immediately, or after processing (removal of overhangs or fill-in by polymerases). MH-mediated joining (right) is facilitated by base-pairing interactions between short stretches of nucleotides at or near the ends of the DSB. (b) MH length in Sμ-Sγ1 (left) and Sμ-Sε (right) junctions from Ku70^−/−HL, Ku70^−/−Lig4^−/−HL, Lig4^−/−HL and WT/HL B cells stimulated in culture with αCD40 plus IL-4. Ku70-, Ku70/Lig4-, and Lig4-deficient cells have more Sμ-Sγ1 (left) and Sμ-Sε (right) joins with longer MH than WT. Data shown are based on 86 or more independent sequences per genotype for Sμ-Sγ1 junctions and 41 or more independent sequences for Sμ-Sε junctions. At least three mice per genotype in three independent experiments were used for Sμ-Sγ1 junctions and one Ku70^−/−HL mouse, two Ku70^−/−Lig4^−/−HL mice, three Lig4^−/−HL, and three WT-HL mice in three independent experiments were used for Sμ-Sε junctions. A detailed summary of all S region junction isolation experiments and mice is presented in Table S4. (C) Percentage of direct Sμ-Sγ1 (left) and Sμ-Sε (right) joins in Ku70^−/−HL, Ku70^−/−Lig4^−/−HL, Lig4^−/−HL, XRCC4^−/−HL and WT/HL B cells. Ku70 and Ku70Lig4-deficient cells have significantly more direct Sμ-Sγ1 and Sμ-Sε joins than Lig4 germline-deficient cells. *, XRCC4 junctions were acquired and published previously (Yan et al., 2007) and are shown here for comparison. Two-tailed Student’s t test was used for p-value. Data shown are based on 86 or more independent sequences per genotype for Sμ-Sγ1 junctions and 41 or more independent sequences for Sμ-Sε junctions. At least three mice per genotype in three independent experiments were used for Sμ-Sγ1 junctions, and one Ku70^−/−HL mouse, two Ku70^−/−Lig4^−/−HL mice, three Lig4^−/−HL, and three WT-HL mice in three independent experiments were used for Sμ-Sε junctions. N, number of independent sequences.