EMSA of normal and AML CD34⁺ cells.

NF-κB binding activity is detected in CD34⁺ AML cells but not in CD34⁺ normal hematopoietic cells by electrophoretic mobility shift assays. (A) Two different normal bone marrow specimens (lanes 1 and 2) and one normal CB specimen (lane 3) were analyzed for NF-κB binding activity in comparison to 6 CD34⁺ AML specimens from patients with different FAB types (specimens 1-6 in Table 1). Nuclear extracts equivalent to 100 000 cells were used for each reaction. (B) Competition assays for 3 of the AML samples from panel A. A 200-fold molar excess of unlabeled oligonucleotide (wild-type or mutant) was used. Supershift assays were done by adding 2 μg per reaction of antibodies against NF-κB subunits p65 or p50. Arrows indicate NF-κB species and asterisks indicate nonspecific bands.