Fig. 2: Spleen and bone marrow isotype-switched memory B cells are distinct in Ig heavy chain repertoire.

a, c Observed overlap between the IgG1/2⁺ or IgA⁺ heavy chain CDR3 repertoire of switched memory B cells from spleen and bone marrow (BM) or b, d random distribution (upper panels). Venn diagrams represent clonotype presence in a given sample: numbers indicate clonotypes present in one organ exclusively or in both (overlap). Random distributions (b, d) show median values for 1000 random distributions drawn while retaining the number of clones initially observed in the sample. Histograms (b, d, lower panels) represent number of clonotypes in spleen and BM in 1000 randomized distributions of observed clonotypes to spleen and BM, dashed red line indicates experimentally observed number of clonotypes present in both spleen and BM. P value of one-sided t-test for difference of randomized overlap against observed. e VH gene recruitment to spleen and BM IgG1/2⁺ (upper panel) and IgA⁺ (lower panel) memory B cells, represented as frequency of a particular VH gene among total CDR3s per organ. Bars show relative abundance of the ten most abundant VH genes, error bars indicate SEM. Significance of difference in VH gene distribution to Spleen and BM assessed by MANOVA, P values corrected for multiple testing (Benjamini-Hochberg), * indicates significant difference in means for a particular VH gene (Welch’s test, two-sided). M1–M3: replicate samples of three female C57BL/6 mice immunized 3× NP-CGG/IFA. Only clones consistently found in technical replicates were considered. Source data for Fig. 2a–e are provided as a Source Data file.