Fig 1: Inversional and deletional recombination of 3'-RSS of DQ52 and DSP2 on WT and IGCR1-mutated IgH alleles.Schematic representation of 3'-RSS of DQ52 (A) and DSP2 (B) rearrangements to VH81X gene segment by inversion (black arrows) or to JHs gene segments by deletion (blue arrows), respectively (left). Products of each form of rearrangements are shown to the right. RAG2-deficient pro–B cell lines with WT or IGCR1-mutated IgH alleles [CBE-/-(1) and CBE-/-(2)] were infected with a Rag2-expressing lentivirus, followed by genomic DNA purification after 14 days of selection with puromycin. LAM-HTGTS experiments were carried out as previously described (33, 39) with baits (red arrows) located 50- to 100-bp upstream of DQ52 (A) or DSP2 (B). Restriction enzyme Sacl-HF (R3156S, NEB) and BseYI (R0635S, NEB) were used to remove germline DNA with DQ52 and DSP2 as bait, respectively. Total reads were aligned to detect recombination by deletion to JHs and by inversion to VH81X. The lower reads of 3' DSP2 RSS utilization compared to DQ52 gene may be due to inefficient restriction of germline DSP2 fragments during library preparation. Average reads and percentages from two independent experiments are shown in red.