RNA-seq analysis of various murine B cell subsets

Purpose: We aimed to link different variations of memory B cell subsets to their proliferating precursors via shared transcriptomic features. Methods : Spleens were processed according to the manuscript. Cells were collected via FACS, washed twice in PBS + 1% FCS, resuspended in 350 ul RLT Plus Buffer (Qiagen) with 1% 2-Me, and frozen at -80 ⁰C. Results: Memory cells have many DEGs in comparison to FO naïve cells. However, differences between memory cells themselves can be attributed to imprinting by their respective proliferating precursors. DP MBCs are split into two major subsets based on transcriptional state. A germinal center B cell program may be required to establish some DEGs in GC-derived memory cells. We used DEGs between DN and DPs, as well as EMPs and GCs, to establish similarity between putative precursors.