Fig. 2.
Fig. 2. Bay 11-7082 specifically inhibits Tax-induced NF-κB activation. / (A) κB-LUC, LTR-LUC, or AP-1-LUC was transfected into JPX-9 cells. JPX-9 cells were pretreated for 1 hour with 5 μM Bay 11-7082 followed by 48 hours of incubation with 20 μM CdCl2 in the continued presence of Bay 11-7082. Relative luciferase activity is expressed relative to the basal level measured in cells transfected with the reporter plasmid without further treatment. To normalize variations, the construct containing the TK promoter-drivenRenilla luciferase (pRL-TK) was cotransfected, and the activities of firefly and Renilla luciferases were measured sequentially from a single sample by means of a dual-luciferase reporter assay system. Data are the mean ± SD of 3 separate transfections. (B) Inhibition of NF-κB/DNA binding in CdCl2-treated JPX-9 cells. Nuclear proteins of JPX-9 cells treated without or with CdCl2 in the absence or presence of Bay 11-7082 were incubated with radiolabeled oligonucleotide containing NF-κB or AP-1 binding site, and DNA binding activity was examined by EMSA.

Bay 11-7082 specifically inhibits Tax-induced NF-κB activation.

(A) κB-LUC, LTR-LUC, or AP-1-LUC was transfected into JPX-9 cells. JPX-9 cells were pretreated for 1 hour with 5 μM Bay 11-7082 followed by 48 hours of incubation with 20 μM CdCl2 in the continued presence of Bay 11-7082. Relative luciferase activity is expressed relative to the basal level measured in cells transfected with the reporter plasmid without further treatment. To normalize variations, the construct containing the TK promoter-drivenRenilla luciferase (pRL-TK) was cotransfected, and the activities of firefly and Renilla luciferases were measured sequentially from a single sample by means of a dual-luciferase reporter assay system. Data are the mean ± SD of 3 separate transfections. (B) Inhibition of NF-κB/DNA binding in CdCl2-treated JPX-9 cells. Nuclear proteins of JPX-9 cells treated without or with CdCl2 in the absence or presence of Bay 11-7082 were incubated with radiolabeled oligonucleotide containing NF-κB or AP-1 binding site, and DNA binding activity was examined by EMSA.

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