Bortezomib activates NF-κB in normal healthy donor-derived PBMCs. (A) PBMCs were cultured with bortezomib (10 nM and 20 nM) for 8 hours. Whole-cell lysates were immunoblotted with indicated Abs. (B) PBMCs and BMSCs from MM patient were cultured with bortezomib (10 nM and 20 nM) for 8 hours. (C) PBMCs were separated into adherent and nonadherent subsets, followed by bortezomib (20 nM) treatment for 8 hours. Nuclear extracts were subjected to EMSA. (D) PBMCs were cultured with or without bortezomib (20 nM for 8 hours). Nuclear extracts were subjected to supershift assays using anti-p65 (65), p50 (50), p52 (52), RelB (B), and cRel (cR) Abs.