Fig. 9
From: PRMT5 is essential for B cell development and germinal center dynamics
Prmt5 acts in the light zone to prevent B cell differentiation. a IHC images for Prmt5 and AID, as GC marker, on consecutive spleen sections from wt mice 14 days post-NP-CGG immunization. Representative of three mice from two experiments. Scale bar, 40 µm. b Representative IF on wt mouse spleen section 10 days after SRBC immunization, stained for GL7 CD35 (LZ marker) and Prmt5. Scale bar, 100 µm. c GSEA of transcriptional changes in Prmt5F/F Cγ1-cre (F/F) versus Cγ1-cre (Ctrl) iGB cells against the indicated gene signatures. Enrichment was considered significant for P < 0.05 and FDR < 5%. d Relative gene expression in positively selected GC B cells. e Proportion of plasma cell-like B cells in Cγ1-cre (Ctrl) or Prmt5F/F Cγ1-cre (F/F) iGB cultures at day 7 post-plating, or at day 3 after replating with 1 ng/mL IL-4 or 10 ng/mL IL-21. Individual mice (symbols) from one experiment are plotted. f Representative flow cytometry histograms of intracellular IgG1 and IgM staining of plasma cells in Prmt5F/F Cγ1-cre (F/F) or Cγ1-cre (Ctrl) mice 6 days post-SRBC immunization. Proportion of IgG1+ and IgM+ plasma cells in individual mice (symbols) from two experiments are plotted. g Representative flow cytometry plots of plasma cells in CD19-cre (Ctrl) or Prmt5F/F CD19-cre (F/F) B cells cultured 4 days in LPS and IL-4. Plasma cell proportion for individual mice (symbols) from three independent experiments are plotted. h Gene expression by RT-qPCR in splenic B cells cultured as in g for 3 days. Mean + s.d. mRNA levels normalized to Actin for n mice from two experiments are plotted relative to Ctrl average. i Mean ± s.d. (or ± s.e.m for n = 2) of plasma cell proportion in splenic B cells stimulated with LPS and IL-4 and treated with various EPZ doses for 4 days, are plotted for n mice from one to five experiments. j Gene expression by RT-qPCR in B cells treated with 5 μM EPZ as in i; individual mice (symbols) are plotted. Bars indicate mean values in all scatter plots. Significant p-values throughout are by an unpaired, two-tailed Student's t test except for GSEA, in which statistics are indicated
