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Blood, Vol. 113, Issue 7, 1464-1473, February 12, 2009
Fc RIIB signals inhibit BLyS signaling and BCR-mediated BLyS receptor up-regulation
Blood Crowley et al.
113: 1464
Supplemental materials for: Crowley et al
Files in this Data Supplement:
- Figure S1. BLyS receptor expression on cultured FO, MZ, and B1 B cells (JPG, 83.2 KB)
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FO, MZ, and B1 B cells were FACS sorted and cultured with the indicated stimuli (concentrations of stimuli are indicated in Methods and Materials). After 24 hours, the cells were harvested and analyzed by flow cytometry for BR3 and TACI expression (BCMA expression was negligible) on live cells. Gray histograms indicate isotype control, unfilled histograms indicate unstimulated levels, and filled black histograms indicate stimulation effect) and median fluorescence intensities (MFI) are shown in bar graphs at bottom. Plots are representative of at least 3 experiments and in all cases n ≥ 3. Significance levels are indicated by p values.
- Figure S2. CD22 cross-linking augments anti-IgM-induced increase in BLyS receptor expression (JPG, 36.1 KB)
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CD23+ B cells were cultured with biotinylated anti-IgM, biotinylated anti-CD22, or both, and cross-linked with avidin (concentrations of stimuli are indicated in Methods and Materials). After 24 hours, the cells were harvested and analyzed by FACS for BR3 and TACI expression on live cells. Histograms are shown at the top (gray histograms indicate isotype control, filled white histograms indicate unstimulated levels, and filled black histograms indicate stimulation effect) and median fluorescence intensities (MFI) are shown in bar graphs at bottom. Plots are representative of 3 independent experiments.
- Figure S3. Mcl-1 and Bcl-2 expression is enhanced by BLyS, but attenuated when combined with BCR ligation (JPG, 23.3 KB)
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Mcl-1 and Bcl-2 message levels, as determined by real-time PCR, following 24 hour culture with the indicated stimuli, with and without the addition of rBLyS at time 0 hour (concentrations of stimuli are indicated in Methods and Materials). Data are representative of 2 independent experiments.
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