Fig. 5: LZ bias in the absence of IL-21 signaling is confirmed by gene expression analysis.

A–C WT and Il21r^−/− mice were immunized i.p. with NP-KLH in alum and on day 7, spleen NP-binding GC B cells (CD138⁻IgM⁻IgD⁻Gr1⁻B220⁺NP⁺IgG1⁺CD38⁻) were sorted and RNA-seq performed. A Mean-difference plot of WT vs Il21r^−/− GC B cells depicts genes that were significantly upregulated in WT compared to Il21r^−/− GC B cells shown in red and significantly downregulated ones in blue (false discovery rate <0.05). B Gene expression fold changes in WT compared to Il21r^−/− GC B cells. Significantly DE genes were ranked by FDR and the 50 lowest then ordered by FC, colored red if upregulated in WT compared to Il21r^−/−, blue if downregulated. C, D limma barcode plots testing for GC zone signature or cMyc signature genes among DE genes from WT vs Il21r^−/− GC B cells as in A. Genes were ranked left to right as most up in Il21r^−/− to most up in WT GC. For zone signature (C), vertical red bars mark DZ signature genes while vertical blue bars mark LZ signature genes. Worms show relative enrichment of DZ and LZ signature genes and camera p values assess significance. The plot shows DZ genes enriched among those up in WT while LZ genes were enriched among those up in Il21r^−/− GC. For the Myc signature (D), vertical red bars mark upregulated genes in Myc⁺ cells while vertical blue bars mark genes upregulated in Myc⁻ cells. Worms show relative enrichment of Myc signature genes and camera p values assess significance. The plot shows Myc^+-associated genes were significantly up in WT GC B cells.