Co-activation of NF-κB and MYC renders cancer cells addicted to IL6 for survival and phenotypic stability

Gene expression profile of Plasmablast without or with MYC, or IKK2 or IKK2 and MYC overexpression B cells from mice were isolated from splenocyte suspensions using CD43 microbeads (Miltenyi Biotec), and plated at a density of 1x106/mL in B-cell media (DMEM high glucose with Glutamax (Gibco), supplemented with Fetal Bovine Serum (GE Healthcare), non-essential aminoacids (Gibco), HEPES buffer (Gibco), sodium pyruvate (Gibco), Penicillin-Streptomycin (Gibco), and 2-Mercaptoethanol (Sigma-Aldrich)). B cells were stimulated with Lipopolysaccharide (LPS; Sigma-Aldrich) at a concentration of 10ug/mL. For gene expression profiling of in vitro derived activated B cells and plasmablasts, cells were FACS sorted at day 3 of LPS stimulation according to the expression of CD19 and CD138 markers (activated B cells – CD19+ CD138neg; plasmablasts – CD19low CD138+).