Figure S2. Panel A shows flow cytometry... | Rockefeller University Press

Figure S2.

Panel A shows flow cytometry plots used to identify different B cell types and plasma cells from the spleens of WT:WT and KO:WT chimeras. Panel B presents bar graphs comparing the total number of marginal zone (MZ) and follicular (FO) mature splenic B cells in WT:WT and KO:WT chimeras. Panel C displays a bar graph quantifying ICOSL surface expression on splenic FO B cells, normalized to the mean MFI of WT CD45.2 positive cells. Panel D includes representative flow cytometry histograms and a bar graph showing TACI surface expression on various cell types from WT:WT and KO:WT chimeras. Panel E shows bar graph comparing the numbers of splenic plasma cells in WT:WT and KO:WT chimeras. Panel F presents bar graph comparing the numbers of splenic T2 B cells in WT:WT and KO:WT chimeras. Panel G shows flow cytometry plots used to identify different B cell types and plasma cells from the spleens of WT:WT and KO:WT chimeras.

Analysis of mixed chimeras. (A) Gating strategy used to identify FO, MZ, T1, T2, and T3 B cells and PCs from the spleens of WT:WT and KO:WT chimeras. (B) Total number of MZ (CD1d^hiIgM^hi) and FO (CD1d^medIgM⁺) mature splenic B cells (CD93⁻B220⁺CD19⁺CD138⁻; CD45.2⁺ and CD45.1⁺ combined) in WT:WT and KO:WT chimeras. Bars show the mean of n = 16 mice, from four independent analyses. Each point represents one mouse. (C) ICOSL surface expression, measured by flow cytometry, on CD45.2⁺ and CD45.1⁺ splenic FO B cells (CD1d^medIgM⁺CD93⁻B220⁺CD19⁺) from WT:WT and KO:WT chimeras, quantified as the geometric MFI and normalized to the mean MFI of WT CD45.2⁺ cells within the same experiment. Bars show the mean of n = 6 mice, from two independent analyses. Each point represents one mouse. Black: WT CD45.2⁺; red: TACI KO CD45.2⁺; gray: WT CD45.1⁺. (D) TACI surface expression, measured by flow cytometry, on TACI KO and WT CD45.2⁺ splenic PCs (TCRβ⁻CD138⁺CD19^lo), MZ (CD93⁻CD1d^hiIgM^hi), FO (CD93⁻CD1d^medIgM⁺), T2 (CD93⁺IgD⁺IgM⁺) and T1 (CD93⁺IgD^lo) B cells (B220⁺CD19⁺), and T cells (TCRβ⁺CD19⁻) from WT:WT and KO:WT chimeras. Left: Representative flow cytometry histograms showing TACI expression on each cell type. Filled: WT; unfilled: TACI KO. Right: TACI expression quantified as the geometric MFI. Bars show the mean of n = 5 mice, representative of two independent analyses. Each point represents one mouse. Black: WT CD45.2⁺; red: TACI KO CD45.2⁺. (E) Numbers of CD45.2⁺ and CD45.1⁺ splenic PCs (TCRβ⁻CD138⁺CD19^lo) in WT:WT and KO:WT chimeras. Bars show the mean of n = 10 mice, from two independent analyses. Each point represents one mouse. Colors are as in C. (F) Numbers of CD45.2⁺ and CD45.1⁺ splenic T2 (CD93⁺IgD⁺IgM⁺) B cells (B220⁺CD19⁺CD138⁻) in WT:WT and KO:WT chimeras. Bars show the mean of n = 16 mice, from four independent analyses. Each point represents one mouse. Colors are as in C. (G) Gating strategy used to identify B6 (Ly9.1⁻) and 129 (Ly9.1⁺) FO B cells, MZ B cells, PCs (CD138⁺B220^loCD19^int-lo), and PBs (CD138⁺B220^hiCD19^int) from the spleens of WT:wt and KO:wt chimeras, for the data shown in Fig. 2. Statistical tests: unpaired t test (B), repeated-measures two-way ANOVA with Fisher’s LSD test (C, E, and F), unpaired Welch’s t test corrected for multiple comparisons using the Holm–Šídák method (D). Numbers above graphs indicate P values where P ≤ 0.05, otherwise no value is shown. PCs, plasma cells; PBs, plasmablasts; MFI, mean fluorescence intensity.