The regulatory role of ZEB2 in ABC formation

To elucidate the direct targets of ZEB2 in ABCs, we performed high-throughput sequencing of regulome by ATAC-seq, CUT & Tag, and CUT & RUN, leading to the identification the accessible sites with ZEB2 binding. Among the genes differentially expressed by Zeb2 deficiency, we found 33 candidate target genes of ZEB2, with 22 repressed and 11 activated by ZEB2. The critical transcription factor Mef2b, essential for GC development, was repressed by ZEB2. This direct regulation was mapped to a conserved region about 20kb downstream of Mef2b's exon I TSS, enriched with enhancer-associated features in both human and mouse. 1.Examination of chromatin accessibility of CD11c+T-bet+ and CD11c−T-bet− B cells sorted by flow cytometry from bm12-induced T-bet reporter mice. 2. Examination of chromatin accessibility of splenic B cells from Zeb2 deficient mice cultured in the presence of mouse ABC-skewing cocktail. 3. Examination of endogenous ZEB2 binding in the in vitro derived B cells. 3. Examination of endogenous ZEB2 binding in the ex vivo sorted B cells.