Figure 3.

Spatial positioning of MBCs

(A–D) Confocal images of (A and D) the lung, (B) mediastinal lymph node, and (C) spleen sections from Aid-EYFP mice treated as in Figure 1A. Scale bars: left 50 μm and right 25 μm in (A), left 300 μm and right 30 μm in (B), left 250 μm and right 55 μm in (C), and 20 μm in (D). The quantification in (A) displays the minimal distance of B cell clusters to the EpCAM⁺ cells. Bar charts in (B) and (C) show the proportion of MBCs residing in the indicated zones. MZ, marginal zone; PALS, periarteriolar lymphoid sheaths.

(E) Dot plot showing CCR6 and CXCR3 fluorescence intensity for MBCs (left). X and Y positions of MBC subsets are laid out in the B220 representation (middle). The quantification displays the distance of each MBC to the cluster’s centroid (right).

(F) Lung images of wild-type and CCR6-deficient Aid-EYFP mice treated as in Figure 1A. Scale bars, 30 μm. The quantification displays the density of YFP⁺ B cells in B cell clusters. Dots represent individual B cell clusters.

(G) Strategy to generate mixed bone marrow chimeras.

(H and I) Histograms showing (H) CCR6 and (I) CXCR3 expression in the lung MBCs and non-B cells in chimeras.

(J and K) Quantification of MBC numbers after 70 days of primary influenza infection in (J) CCR6- or (K) CXCR3-deficient chimeras.

(L and M) Enumeration of ASCs after 4 days of re-challenge in (L) CCR6- or (M) CXCR3-deficient chimeras. The quantification of one representative experiment is shown in the bar charts; each dot represents one mouse. In all panels: mean ± SEM, t test: ^∗∗p < 0.01, ns, not significant.

See also Figure S3.