![Figure 9 MZCD9+ B-cell subset is poised for innate immune responses. Heatmap display of NGS data showing enrichment of (A) TLR and (D) innate immune sensors/effector genes. Asterisks denote important genes discussed in the text. qRT-PCR analysis validated NGS results showing MZCD9+ express highest amounts of TLR3, TLR7 and Perforin-2. qRT-PCR (black bars) and RNA-Seq (gray bars) of (B) TLR3, (C) TLR7, and (E) Perforin-2. qRT-PCR data are representative of three independent experiments with FACS-sorted B-cell subsets, where each experiment utilized pooled splenocytes from 2 to 5 mice. (F) qRT-PCR of Perforin-2 mRNA fold-change after stimulation with endosomal TLR agonists. B220 positively selected cells (>95% purity) were stimulated 18 h with either TLR3 agonist Poly(I:C), TLR7/8 agonist CL097, or TLR9 agonist CpG at 1 μg/mL and compared to NS. (G) qRT-PCR of Perforin-2 mRNA fold-change using B220 positive (>95% purity) or CD43 negative (~90% purity) B-cells stimulated with Poly(I:C) as in (F). Data are representative of four independent experiments. (H) qRT-PCR of Perforin-2 mRNA fold-change using FACS derived MZCD9+ or FO-I B-cells treated with Poly(I:C) as in (F). *P ≤ 0.05; **P ≤ 0.01.](https://i0.wp.com/idsc.miami.edu/wp-content/uploads/2020/10/fimmu-06-00030-g009.jpg?resize=730%2C350&ssl=1)
Distinct Transcriptomic Features and B-Cell Populations in the Mouse…
Splenic transitional B-cells (T1 and T2) are selected to avoid self-reactivity and to safeguard against autoimmunity, then differentiate into mature follicular (FO-I and FO-II) and marginal zone (MZ) subsets. Transcriptomic analysis by RNA-seq of the five B-cell subsets revealed T1 cell signature genes included RAG suggesting a potential for receptor revision. Read more “Distinct Transcriptomic Features and B-Cell Populations in the Mouse Spleen”