Figure 5

Partial purification of NF-κB-inducing factor derived from MSCs. (A) EMSA analysis of RPMI8226 cells cultured alone or with H1-ESC-MSCs (ES-MSCs) for 24 hours with or without 4-hour treatment of 100 nM bortezomib (Bort). (B) EMSA analysis of RPMI8226 cells cultured alone or with CM from H1-ESC-MSCs (CM) for 24 hours with or without 4-hour treatment with 100 nM bortezomib (Bort). NF-κB dimers and fold-change in DNA-binding activity are displayed as before. (C) EMSA analysis of RPMI8226 cells untreated (OT) or treated with 1× CM or sequential fractions eluted from anion-exchange column with increasing NaCl concentration gradient (Fraction 1-8). (D) EMSA analysis of RPMI8226 cells untreated or treated with increasing aliquots of an activity-containing fraction eluted from the anion-exchange column (Fr3), with or without treatment with 100 nM bortezomib (Bort). (E) EMSA analysis of RPMI8226 cells treated with 100 nM bortezomib (bort), 10 ng/mL rhTNF-α (TNF-α) or both.