Research Paper Volume 8, Issue 12 pp 3321—3340

Figure 5. Molecular targeting of NF-κB in diffuse large B-cell lymphoma (DLBCL) cell lines. (A) DLBCL-MS cells were transfected with empty control vector or a pCMV-ΙκBαM vector for 24 hrs. Nuclear extracts (10 μg) were analyzed for NF-κB expression by EMSA. Cytoplasmic extracts were assessed for ΙκBα and actin protein expression by Western blotting. (B) Transfected cells from part A were also assessed for apoptosis after 24 hours of incubation using annexin V assays. (C) MS cells were transfected with plasmids expressing the p52, RelB, p65, c-Rel, or a non-specific (NS) shRNA. Forty-eight hours post-transfection, proteins were extracted and analyzed for NF-κB component inhibition by Western blot. (D) Indicated DLBCL cell lines were transfected with the validated green fluorescent protein (GFP)-plasmid–based shRNA for each of the NF-κB subunits. After 16 hours, GFP–positive cells were sorted and assessed using proliferation assays. Data represent two independent experiments with triplicate samples. Abbreviations: GCB, germinal center B-cell–like; ABC, activated B-cell–like, DN, dominant negative.