VDJ analysis of IL-10+ and IL-10- splenic B1a, MZ, and FO B cells from sIgM-/- mouse

We compared the clonotypes of IL-10+ and IL-10- B1a, marginal zone and follicular B cells from the spleen of sIgM-/-IL10GFP mouse using 10X single cell RNAseq to analyze potential differences in clonality between IL-10+ and IL-10- B cells. Overall design: splenic B cells from sIgM-/-IL10GFP mice were stimulated for 4 hours with PMA/Ionomycin/LPS and processed for CD19 magnetic bead enrichment followed by FACS sorting for B1a, marginal zone, and follicular B cell populations. IL-10+ and IL-10- B cells from each subset were sorted separately based on GFP expression or absence.