Fig. 4

General repertoire features of appendix and tonsil B cell subsets. Donors and samples used for this analysis are shown in Tables S1 and S2 and exclude the 12-hr-old donor. N = 7 (A) Clone counts for each tissue and subset aggregated across donors (B) Fraction of sequence copies comprising the top 20 clones (D20 Index) for each tissue and subset (C) VH-gene usage by tissue/subset combination normalized by row. Each clone is counted only once per subset/tissue combination. White boxes indicate no data. VH genes occurring with a frequency of < 1% in all samples are excluded and V-ties for VH3-23 were combined. The frequency of VH4-34 (*) was significantly increased in IgM + IgD + compared to IgM + IgD- and class-switched (p < 0.001 by one-sided binomial test) (D) Average clonal CDR3 nucleotide length in each tissue and subset combination. Each dot indicates an individual. (E) Fraction of clones with at least 2% mutation in the VH region. Each dot indicates an individual. (F) Clonal mutation percentage. Each dot indicates an individual. Panels D-F show averages per person, but statistical tests were performed on the underlying data: differences between subsets are all significant (p < 0.0001) and all tissue comparisons within the same subset are significant (p < 0.001) except the CDR3 length for IgM + IgD+ (Mann-Whitney U-test). (G) Distribution of SHM for each tissue stratified by subset. The x-axis shows VH gene SHM percentage, the y-axis shows the fraction of clones with the given SHM level, and each trend line indicates subset. NT = nucleotide. Gating strategy for cell sorting (Supplementary Fig. 5).