Fig. 1

Regulated Prmt5 expression in B cells. a Prmt5 transcript levels in B cell stages from the indicated datasets, each normalized to follicular (Fo) B cells. RT-qPCR data were normalized to Actin mRNA and obtained from a pool of two mice sorted for Hardy’s BM fractions (FrA to E), or from splenic B cells from two mice stimulated ex vivo with LPS and IL-4 for 48 h (Activated). CLP common lymphoid progenitor, T1–T3 transitional B cells, MZ marginal zone B cells, PC plasma cells, Sp spleen. b Prmt5 expression kinetics and sDMA-modified proteins by WB in extracts of WT splenic B cells stimulated with LPS (5 µg/mL) and IL-4 (5 ng/mL) probed with anti-PRMT5, -Actin (as loading control), and -sDMA (SYM11) antibodies. c Absolute number of B cell subpopulations in the spleen of CD19-cre (Ctrl) and Prmt5^F/F CD19-cre (F/F) mice. Individual mice (dots) and mean (bars) values are plotted. Gatings in Supplementary Fig. 1B. d Representative WB of resting splenic B cell extracts from CD19-cre (Ctrl) and Prmt5^F/F CD19-cre (F/F) mice. Means + s.d. levels of Prmt5 normalized to Actin quantified from n mice by WB are plotted relative to the Ctrl. e Representative Prmt5, Actin, and sDMA (SYM11) WB and quantitation as in d, in extracts of splenic B cells activated with LPS (5 µg/mL) and IL-4 (5 ng/mL) for 72 h. f Prmt5 mRNA (by RT-qPCR) as a function of Prmt5 protein (by WB) at 72 h post-stimulation for individual mice. Spearman’s test correlation coefficient (r) and p-value (p) are indicated. g Representative immunohistochemical staining for PNA as GC marker and Prmt5 on consecutive mouse spleen sections at day 14 post-immunization. Scale bars, 500 µm