Panel A: Flow cytometry plots show the gating strategy for analyzing B cell development in the bone marrow of WT, M-only, and mM-only mice. The plots depict various stages of B cell maturation, with axes labeled for different cell surface markers. Panel B: Flow cytometry plots illustrate the gating strategy for analyzing B cell development in the spleen, showing different B cell subsets with labeled axes for specific markers. Panel C: Flow cytometry plots display the gating strategy for analyzing B cell development in the mesenteric lymph nodes (mLN), with axes labeled for various cell surface markers. Panel D: Flow cytometry plots show the gating strategy for analyzing B cell development in the Peyer's patches (PP), with axes labeled for different markers. Panel E: A scatter plot shows the percentages of different splenic B cell subsets among live Dump- cells, with the x-axis labeled for B cell types and the y-axis for percentages. Panel F: A scatter plot depicts the percentages of transitional T1 and T2 B cells in live Dump- cells, with the x-axis labeled for B cell types and the y-axis for percentages. Panel G: A scatter plot shows the percentages of splenic B1a, B1b, and B2 cells among live Dump- cells, with the x-axis labeled for B cell types and the y-axis for percentages. Panel H: A scatter plot illustrates the percentages of splenic light chain bearing mature B cells, with the x-axis labeled for light chain types and the y-axis for percentages. Panel I: A bar graph shows the usage of Iglv genes in follicular B cells, with the x-axis labeled for gene names and the y-axis for frequencies. Panel J: A scatter plot depicts the geometric mean fluorescence intensity (GMFI) of IgM-FITC in bone marrow mature B cells and splenic follicular B cells, with the x-axis labeled for cell types and the y-axis for GMFI values. Panel K: A bar graph shows the proportions of Dark and Light Zone B cells in mesenteric lymph nodes at steady state, with the x-axis labeled for zone types and the y-axis for proportions. Panel L: A scatter plot depicts the proportions of T follicular helper cells in mLN at steady state in total CD3 positive T cells, with the x-axis labeled for mouse strains and the y-axis for proportions.
Analysis of M-only and mM-only mouse strains. Related to Fig. 1. (A) Flow cytometric gating strategy in the bone marrow. All stainings were cell surface and not intracellular stainings. (B) Flow cytometric gating strategy in the spleen. (C) Flow cytometric gating strategy in the mLN. (D) Flow cytometric gating strategy in the PP. (E) Percentages of splenic, transitional (Transit), pre-marginal zone (pre-MZB), marginal zone (MZB), and follicular (FoB) cells among live Dump− cells. (F) Percentages of transitional T1 and T2 B cells in live Dump− cells. (G) Percentages of splenic B1a, B1b, and B2 cells among live Dump− cells. (H) Percentages of splenic κ or λ light chain–bearing mature B cells. (I) Bar graphs showing Iglv gene usage in follicular B cells. Top 10 most frequent genes ranked by the mean frequency in mM-only mice are shown. (J) GMFI of IgM-FITC in the bone marrow mature B cells and splenic follicular B cells. (K) Proportions of dark and light zone B cells in mLNs at steady state. (L) Proportions of T follicular helper cells in mLN at steady state in total CD3+ T cells. Each dot represents a single mouse. *P > 0.05. Bars indicate the mean ± SD. PP, Peyer’s patches; GMFI, geometric mean fluorescence intensity. DZ, dark zone; LZ, light zone.