FIGURE 2.

Binding of the minimal promoter fragment and differential expression patterns of IRF and NFAT. A, ChIP assays were performed on 293T and CEM-A cell lysates using antibodies against NFATc1, NFATc2, or IRF-4. Pulldown was detected by quantitative real-time PCR and percent of input determined by comparison with an Ig control pulldown. Average cycle threshold of GAPDH for input samples was used as an internal control for quality of input DNA, 293T Ct 29.0 ± 1.47, CEM-A Ct 28.6 ± .86. B, 293T, HeLa, and CEM-A cell lysates were subcellularly fractionated and probed for the presence of NFAT and IRF family members. Immunoblots probed with an antibody specific for actin served to monitor loading equivalency. Ct, threshold cycle.