Identification of B cell stimulation-specific A20-repressed genesets

Loss of one copy of the negative feedback regulator A20 is implicated in the development of autoimmunity. In mouse models the B cell specific loss of A20 can induce autoimmune phenotypes. To identify genesets which are repressed by A20 in B cell, we stimulated A20-deficient and control B cells in vitro with αIgM, αCD40 or LPS for 3 or 14 hours to reflect B cell receptor engagement, T cell help and innate/TLR signals, respectively, and analyzed the effect on gene expression. Per biological replicate, splenocytes from three mice were pooled and follicular B cells were enriched by depletion of CD9+ (marginal zone B) and CD43+ (T) cell. 7 million cells were stimulated with anti-IgM, anti-CD40 or LPS or not stimulated (control) for 3 hours or 14 hours.