Extended Data Figure 4: Orientation-biased joining between AID-initiated rSμ and downstream AID-initiated S-region breaks in anti-CD40/IL4-activated and LPS-activated Sμ-truncated B cells.
From: Orientation-specific joining of AID-initiated DNA breaks promotes antibody class switching
a, 150-bp rSμ sequence used as HTGTS bait with red arrow indicating rSμ 5′-broken end HTGTS primer; red and blue vertical lines indicate canonical AGCT or other RGYW AID-targeting motifs, respectively. Distribution of rSμ break points in junctions to downstream S regions recovered from anti-CD40/IL4-stimulated rSμ B cells. b, HTGTS analyses of anti-CD40/IL4-activated rSμ B cells, 5′ rSμ AID-initiated broken end (red primer, n = 3) junctions to AID-initiated DSBs in Sγ1 and Sε which includes deletion (− orientation, red) or inversions (+ orientation, blue). c, HTGTS analyses of anti-CD40/IL4-activated rSμ B cells, 3′ rSμ AID-initiated broken end (blue primer, n = 3) junctions to AID-initiated DSBs in Sγ1 and Sε which includes excision circle (+ orientation, blue) or inversions (− orientation, red). d, Bar graph showing the percentage of junctions (average ± s.d.) from anti-CD40/IL4-activated rSμ 5′-broken end libraries mapped to Sγ1 and Sε. e, Bar graph showing the percentage of junctions (average ± s.d.) from anti-CD40/IL4-activated rSμ 3′-broken end libraries mapped to Sγ1 and Sε. f, HTGTS analyses of LPS-activated rSμ B cells, 5′ rSμ (red primer, n = 3) AID-initiated broken end junctions to AID-initiated DSBs in Sγ3, Sγ2b and Sγ2a which include deletional joining (− orientation, red) or inversions (+ orientation, blue). g, HTGTS analyses of LPS-activated rSμ B cells, 3′ rSμ (blue primer, n = 3) AID-initiated broken end junctions to AID-initiated DSBs of above LPS-stimulated cells in Sγ3, Sγ2b and Sγ2a which include excision circle (+ orientation, blue) or inversions (− orientation, red). h, i, Percentage of junction distribution at Sγ3, Sγ2b and Sγ2a in both orientations from both 5′-broken end libraries (h) and 3′-broken end libraries (i) are shown as average ± s.d. from three independent experiments. For detailed legends and further discussion refer to the Supplementary Information.
