Fig. 6
From: PRMT5 is essential for B cell development and germinal center dynamics
Prmt5 regulates gene expression and maintains splicing fidelity in B cells. a Volcano plot of gene expression changes in Prmt5F/F Cγ1-cre versus Cγ1-cre iGB cells. The number of genes significantly changed by ≥2- or ≥4-fold (in red), are indicated. The violin plot shows the absolute change for the genes changing by ≥2-fold, p value from two-sided t-test. b GO terms significantly enriched for the 257 genes upregulated ≥2-fold in Prmt5-null iGBs, analyzed against a background list of all expressed genes in iGBs at the DAVID server. c GSEA of a curated set of 346 genes that are direct targets of p53 and are upregulated in response to p53. d Venn diagrams comparing genes differentially expressed by ≥1.5-fold to genes with at least one splicing alteration. Only significant events (P-value <0.05) for well-expressed genes (basemean >50) are included. e Scatter plot of inclusion level difference (ILD) in WT/KO for each significant splicing event (P < 0.05, ≥10% ILD) separated by category (SE, skipped exon; MXE, mutually exclusive exons; A5SS and A3SS, altered 5′ and 3′ splicing site; RI, retained intron). Violin plots for SE and RI highlight the preferential direction of the change in each. f Venn diagram of significant skipped exon (SE) and retained intron (RI) events. g Sequence logos of the splicing donor and acceptor sites around RI and SE events in Prmt5F/F Cγ1-cre iGBs. Each letter’s height represents the probability of appearing at that position. h Venn diagrams of chosen overlapping GO terms containing genes with significantly affected splicing in Prmt5-null B cells. Selected genes involved in DNA repair are indicated along with ILD and splicing event type. i Representative histograms of γH2AX staining in Cγ1-cre (Ctrl) and Prmt5F/F Cγ1-cre (F/F) mice GC B cells (B220+ Gl7+ CD95+), 10 days post-SRBC immunization. Proportion of γH2AX+ and normalized γH2AX levels estimated from mean fluorescent intensity (MFI) are plotted for five mice (symbols) per genotype from two experiments. p-Values are by an unpaired, two-tailed Student's t test
