Figure 7.
Inhibition of NF-κB–mediated signaling by twist1 is promoter-specific. DO11.10 Th cells were stimulated with OVA327-339, APCs, and 1 ng/ml IL-12. On d 2, cells were infected with control virus, twist1, or I-κBαM-encoding virus. On d 3, cells were nucleoporated with a mixture of a plasmid encoding Renilla luciferase, and a firefly luciferase reporter construct, driven by a NF-κB–responsive promoter (4xκB-luc). (A) Cells were restimulated with PMA/ionomycin for 6 h, sorted according to expression of the viral marker gene gfp, and luciferase signals were quantified in duplicates (mean ± SD). (B) The very same Th1 cultures were restimulated on d 5 and stained for intracellular cytokine expression. Frequencies of cytokine-expressing cells among infected, i.e., GFP+CD4+ T cells, are displayed. Data are representative of two experiments.