Figure 5
A, Protein components of protein-DNA complexes formed between the nuclear extracts from RA synovial cells and a NF-kB oligonucleotide probe. Synovial cells were treated with (lane 2 to lane 7) or without IL-1β (lane 1). After incubation with a 32P-labeled NF-κB oligonucleotide probe, samples were run on 4% native PAGE and gel was dried and autoradiographed. If indicated, 20 times molar excess of competitor oligonucleotides or antibodies were included in the reaction mixtures. C1, C2, and C3, positions of protein-DNA complexes were formed between the NF-kB probe and nuclear extracts. F, position of free probe. B, Effect of IL-1β, Dex, and FK506 on NF-κB DNA-binding activity. Synovial cells were treated with IL-1β, Dex, and FK506, as indicated, and nuclear extracts were prepared and C2 complex formation was monitored in EMSA. C, Effect of IL-1β, Dex, and FK506 on NF-κB DNA-binding activity. NF-κB DNA-binding activity (C2 position) was calculated as a relative unit by densitometer. IL-1β control was assigned the value of 100%, and data were expressed as the percentage of IL-1β control. Data represent the mean ± sd of two separate experiments.
