Fig. 4.
Diversification of SCLs. a Diversification of VH and VL sequences in TSA-treated L15H15 cells. Two independent clones (B7-9 and B7-11) were treated for 90 days with or without TSA. The numbers of unique sequences (NUS) in 239 randomly sampled sequences of the VH (left) and VL (right) were examined on days 0, 21, 42, and 90. b Schematic VH sequences in L15H15 cells cultured with TSA for 42 days. Horizontal lines represent the sequence reads, and the numbers of identical sequences in the observed whole sequences are shown to the left of the horizontal lines. Open boxes represent putative GC tracts, and the corresponding putative pseudogenes used as templates of GC are described in the abbreviated name (for example, pseudogene “VH08” is described as “08”). Closed ovals show single-nucleotide substitutions that have corresponding sequences in pseudogenes, whereas open ovals are single-nucleotide substitutions that are not attributed to GC. Closed and open triangles represent deletions and insertions, respectively. c Comparison of the diversities of L15H15- and L30H45-derived libraries. NUS in 500 randomly selected VH (left) and VL (right) sequences from L15H15-derived (open and closed circles) and L30H45-derived (open and clone triangles) libraries at day 21 (closed circles and triangles) or day 42 (open circles and triangles) of TSA treatment (n = 6 for L15H15-derived libraries and n = 12 (left) and n = 11 (right) for L30H45-derived libraries. Long horizontal bars represent the mean, and their values are shown alongside. Error bars represent ± s.d.). *P < 0.05, **P < 0.01, and ***P < 0.001