Genome-wide analysis of pSTAT3 (Tyr705) in diffuse large B-cell lymphoma

Diffuse large B cell lymphoma (DLBCL), the most common non-Hodgkin lymphoma, includes two main molecular subtypes: activated B cell-like (ABC) DLBCL and germinal center B cell-like (GCB) DLBCL. ABC DLBCL is more aggressive, with the activated JAK1-STAT3 pathway that promotes cell survival. In order to study the underlying pathogenic mechanism, we performed a ChIP-seq experiment by phospho-STAT3(Tyr705) antibody in ABC DLBCL cell lines, and identified 3,456 STAT3 target genes genome-wide. In parallel, ChIP-seq experiment was performed with STAT3 antibody in a-IgM stimulated naïve B cells for verification of these targets of STAT3 protein. TMD8 was treated by eithor DMSO or AZD1480 (JAK inhibitor) for 4hrs, ChIP-seq experiments were performed by pSTAT3(Tyr705) antibody (#9145S, Cell signaling). Naïve B cells were collected from peripheral blood mononuclear cells by Naive B Cell Isolation kit (miltenyi biotec; 130-091-150), and treated by 10ug/ml a-IgM (southernbiotech; #2020-01) for 24hrs. ChIP-seq experiments were performed by STAT3 antibody (SC-482X, Santa Cruz). Overall design: TMD8 cells were treated by eithor DMSO or AZD1480(JAK inhibitor) for 4hrs. Naïve B cells were collected from peripheral blood mononuclear cells by Naive B Cell Isolation kit (miltenyi biotec; 130-091-150), and treated by 10ug/ml a-IgM (southernbiotech; #2020-01) for 24hrs.