Figure 7

Effects of anti-CD3 plus anti-CD28 on NF-κB complex in nuclear extracts of rheumatoid arthritis (RA) peripheral blood mononuclear cells (PBMC) and normal PBMC. (a) NF-κB activity in the absence (lane 1) or presence (lane 2) of anti-CD3 plus anti-CD28 antibody; supershift assay of NF-κB site with antibodies against p65 (lane 3), p50 (lane 4) and c-Rel (lane 5). PBMC from patients with RA were stimulated with anti-CD3 plus anti-CD28 and were used for the supershift assay. (b) NF-κB activity in the absence (lane 1) or presence (lane 2) of anti-CD3 antibody plus anti-CD28 in normal PBMC. (c) NF-κB activity in the absence (lane 1) or presence (lane 2) of anti-CD3, anti-CD3 plus LY294002 (lane 3) or anti-CD3 plus pyrrolidine dithiocarbamate (PDTC; lane 4). Arrows denote a labeled oligonucleotide band shifted after NF-κB binding. The lower panel shows an immunoblot for IκB-α and actin at the same time.