FIG 2.

(A) Flow chart summarizing the high-throughput screen for direct inhibitors of p65 dimerization. (B) Chemical structure of WFA. (C) Dose-response to WFA of the empty plasmid and p65-N/C-RL. (D) Cells were cotransfected with p65 and GFP-p65 and harvested 48 h later. The cell lysates were immunoprecipitated with anti-GFP and treated with 10 μM or with 30 μM WFA for 1 h. The immune complexes were washed and then analyzed by Western blotting using anti-p65 antibody. (E) The N and C termini of the RL fused to p65 or to p50 were transfected into cells, and RL activity was measured after 24 h. The bars represent the means ± SEM of the results of 3 independent experiments. The activity of the WT p65-C/N-RL pair was set to 1. (F) Dose-response to WFA of the empty plasmid, a homodimer of p50-N/C-RL, and a heterodimer of p65-C/p50-N-RL. The asterisks denote statistically significance differences (P < 0.05).