Changes in NFs during TNF-α–induced maturation of MDDC.

(A) Recognition of NF-κB, C/EBP, and AP-1 consensus sequences by nuclear transcription factors from immature (I) and mature (M; treated with TNF-α for 96 hours) MDDC. EMSA was done on the corresponding consensus oligonucleotide probes by using nuclear extracts and in the absence (lanes marked with a minus sign) or presence (lanes marked with a plus sign) of unlabeled competitor (Comp) oligonucleotides at a 100-fold molar excess. (B) Determination of NF-κB DNA-binding activity in total cell extracts from LPS-treated or TNF-α–treated MDDC cultured for 24 or 48 hours in the absence or presence of the indicated inhibitors (U0 indicates U0126; and SB, SB203580). Competition was done by adding cold NF-κB consensus oligonucleotide at a 100-fold molar excess.