Skip to main content
. 2014 Nov 6;33(24):2922–2936. doi: 10.15252/embj.201490332

Figure 4. RTCB is required for immunoglobulin secretion by antibody-secreting cells in vitro.

Figure 4

B220+ cells were enriched from the spleen of control (Rtcbfl/fl or Rtcbfl/+), Rtcbfl/+ Cd23-Cre or Rtcbfl/fl Cd23-Cre mice and cultured in the presence of 20 μg/ml LPS for 3 days.
  1. IgM ELISA. Identical numbers of FACS-sorted CD138+ CD22− plasmablasts or CD138− CD22low pre-plasmablasts of the indicated genotypes were plated for 24 h prior to ELISA analysis of their supernatants. The data are presented as relative units (RU) compared to control cells (n = 4, mean and SEM are displayed). An unpaired Student's t-test was used to analyze the statistical significance (**P < 0.01, ***P < 0.001, ****P < 0.0001).
  2. IgM ELISPOT analysis. FACS-sorted CD138+ CD22− plasmablasts (500 cells) were plated for 16–18 h. A representative assay is shown in the top panel. Bar diagrams in the low panel show the average number of IgM-secreting cells and their median spot size (measured in pixels), respectively (n = 4 mean and SEM are displayed). An unpaired Student's t-test was used to analyze the statistical significance (*P < 0.05, **P < 0.01).
  3. Plasmablasts were analyzed by electron microscopy. Representative plasmablasts of the indicated genotypes are shown.