SFRs suppress TCR signal strength in Tfh cells and promote expression of pro-survival effectors in GC B cells. (A and B) WT or SFR KO mice were bred with mice expressing GFP under the control of the Nur77-encoding promoter. Mice were then analyzed as detailed in A. GFP expression in total B cells, GC B cells, total CD4+ cells, Tfh cells, and GC Tfh cells was assessed by flow cytometry (B). Red histograms, Nur77-GFP− control mice; blue histograms, Nur77-GFP+ mice (WT, n = 11; SFR KO, n = 12). Data are pooled from three independent experiments. (C) GC B cells expressing activated caspases were detected by staining with FITC-VAD-FMK (n = 5 mice per genotype). Control was with addition of Z-VAD-FMK. Data are pooled from two independent experiments. (D and E) Mice were immunized as detailed in Fig. 2 B. Expression of IgLCs and Bcl-2 (D) and Bcl-2 and Bcl-xL (E) was determined at days 7 and 8. The mean fluorescence intensities (MFIs) of IgLCs in total GC B cells and Bcl-2hiIgLChi GC B cells were determined (D). DL/DH, ratio of the number of Bcl-2loIgLClo cells to the number of Bcl-2hiIgLChi cells. The MFIs of Bcl-2 and Bcl-xL in total GC B cells were determined (E). DL/DH, ratio of the number of Bcl-2loBcl-xLint cells to the number of Bcl-2hiBcl-xLhi cells. As control, cells were stained with isotype controls. In D, WT, n = 3; SFR KO, n = 3; data are from one experiment representative of three experiments (day 7); n = 3; SFR KO, n = 4; data are from one experiment (day 8). In E, WT, n = 9; SFR KO, n = 10 (day 7); WT, n = 6; SFR KO, n = 7; data are pooled from three (day 7) or two (day 8) independent experiments. Each symbol represents a mouse. *, P < 0.05; **, P < 0.01; ***, P < 0.001 (two-tailed Student’s t test). Data are shown as mean ± SEM. D, day; ns, not significant.
Figure 8.

SFRs suppress TCR signal strength in Tfh cells and promote expression of pro-survival effectors in GC B cells. (A and B) WT or SFR KO mice were bred with mice expressing GFP under the control of the Nur77-encoding promoter. Mice were then analyzed as detailed in A. GFP expression in total B cells, GC B cells, total CD4+ cells, Tfh cells, and GC Tfh cells was assessed by flow cytometry (B). Red histograms, Nur77-GFP− control mice; blue histograms, Nur77-GFP+ mice (WT, n = 11; SFR KO, n = 12). Data are pooled from three independent experiments. (C) GC B cells expressing activated caspases were detected by staining with FITC-VAD-FMK (n = 5 mice per genotype). Control was with addition of Z-VAD-FMK. Data are pooled from two independent experiments. (D and E) Mice were immunized as detailed in Fig. 2 B. Expression of IgLCs and Bcl-2 (D) and Bcl-2 and Bcl-xL (E) was determined at days 7 and 8. The mean fluorescence intensities (MFIs) of IgLCs in total GC B cells and Bcl-2hiIgLChi GC B cells were determined (D). DL/DH, ratio of the number of Bcl-2loIgLClo cells to the number of Bcl-2hiIgLChi cells. The MFIs of Bcl-2 and Bcl-xL in total GC B cells were determined (E). DL/DH, ratio of the number of Bcl-2loBcl-xLint cells to the number of Bcl-2hiBcl-xLhi cells. As control, cells were stained with isotype controls. In D, WT, n = 3; SFR KO, n = 3; data are from one experiment representative of three experiments (day 7); n = 3; SFR KO, n = 4; data are from one experiment (day 8). In E, WT, n = 9; SFR KO, n = 10 (day 7); WT, n = 6; SFR KO, n = 7; data are pooled from three (day 7) or two (day 8) independent experiments. Each symbol represents a mouse. *, P < 0.05; **, P < 0.01; ***, P < 0.001 (two-tailed Student’s t test). Data are shown as mean ± SEM. D, day; ns, not significant.

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