Western blot analysis of extracts from Vero cells, untreated or treated with TNF-α #2169 (20 ng/ml) for the times indicated, using Phospho-NF-κB p105 (Ser933) (18E6) Rabbit mAb #4806 (upper) and NF-κB p105/p50 Antibody #3035 (lower).
Western blot analysis of extracts from Hela cells, untreated or treated with TNF-alpha (12ng/ml) for the indicated amounts of time, using NF-kappaB p105/p50 Antibody.
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 HeLa cells treated with Human Tumor Necrosis Factor-α (hTNF-α) #8902 (30ng/ml) for 1 hour and either 20 μl of NF-κB1 p105/p50 Antibody #3035 or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP ® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by Real-Time PCR using SimpleChIP ® Human IkB-alpha Promoter Primers #5552, human IAP2 promoter primers, and SimpleChIP ® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.