AG-490 fails to inhibit TNF-–induced NF-κB DNA binding.

D10 cells were treated with the DMSO control (lanes A-E) or 100 μmol/L AG-490 (lanes F-J) and incubated with medium (−) or 20 ng/mL TNF-α (+) for 10 minutes at 37°C. Nuclear extracts corresponding to 5 μg of protein were incubated in the absence (lanes A, B, F, and G) or presence of antibody α-NF-κB p50 (lanes C and H), p65 (lanes D and I), or normal rabbit serum (nrs; lanes E and J) and then with a ³²P-labeled oligonucleotide probe. Arrow indicates migrational location of each nonsupershifted NF-κB–DNA complex or free probe.