Fig. 6

IgM⁺IgD⁻ clonal overlap analysis. (A) CDR3 AA overlap in each subset (see methods for filtering conditions) colored by IgM/IgD overlap. Each row indicates a subset, and each column represents a unique CDR3 AA sequence. Dark red indicates clonal presence in both IgM⁺IgD⁻CD27⁺ and IgM⁺IgD⁻CD27⁻, bright red indicates presence in IgM⁺IgD⁻CD27⁺ but not IgM⁺IgD⁻CD27⁻, and pink indicates presence in IgM⁺IgD⁻CD27⁻ but not in IgM⁺ IgD⁻ CD27⁺. Gray indicates lack of presence in either IgM⁺IgD⁻ subset. Only sequences that overlap in at least two subsets are shown. (B) Venn diagrams showing the numbers of clones in IgM⁺IgD⁺, IgM⁺IgD⁻ and class switched subsets. The bold numbers and dashed lines indicate the group of class-switched clones that overlap with IgM⁺IgD⁻ from which lineages in panel C were selected. (C) Three selected lineages from different donors showing individual clones that have IgM⁺IgD⁻ and IgA⁺ sequence variants. Nodes are colored by subset. Black nodes are inferred. Numbers under the nodes indicate the number of somatic mutations in the VH gene relative to the preceding vertical node. GL = germline. Data are limited to individuals with sorted subsets (Supplemental Table S3) and exclude the 12-hr old donor. Gating strategy for cell sorting (Supplementary Fig. 5).