Comparison of NF-κB activity between ALCL and HD.
(A) NF-κB DNA binding activity of ALCL (lanes 1 to 4) and HD (lanes 5 to 7) cells was analyzed by EMSA. The whole-cell lysates were incubated with 32P-labeled oligonucleotide probe containing a κB site and resolved on a 5% nondenaturing polyacrylamide gel. Supershift assay of Karpas 299 cells using antibodies against p65, p50, and p52 determined the components of NF-κB binding complexes (lanes 11 to 13). Lane 9, addition of an excess amount of unlabeled probe; lane 10, addition of the unlabeled AP2 probe (5′-GATCGAACTGACCGCCCGCGGCCCGT-3′) to eliminate nonspecific binding. (B) Western blotting analysis of ALCL and HD cell lines for the expression of p65, p50, and p52. Cell lysates were prepared from 1 × 106 cells, and an aliquot was loaded.
