EZH2-dependent chromatin accessibility changes during B cell differentiation

To understand the role of EZH2 in B cell differentiation, EZH2 was inducibly deleted using tamoxifen and B cells stimulated to differentiate with LPS in vivo. After 3 days, EZH2-sufficient and EZH2-deficient naive B cells and plasmablasts were FACS isolated from the spleens and ATAC-seq was performed to identify the chromatin accessibility changes that are programmed by EZH2. Bone marrow chimeric mice were established from CD45.1/2 CRE negative and CD45.2 CRE positive Ezh2-floxed mice to study the cell intrinsic role of Ezh2 during B cell differentiaiton. Chimeric mice were treated with tamoxifen to induce deletion of Ezh2 in CD45.2 cells and subsequently inocculated with LPS to induce differentiation. Three-days later naïve B cells (nB) and differentiated plasmablasts (Pb) were isolated from Ezh2-sufficient (CD45.1/2) and Ezh2-deficient (CD45.2) populations by FACS.