text: Oligonucleotides (Oligos) and 5'-32-P-Labeling
  The complementary double-stranded (ds) oligonucleotides (oligos) from the TNFalpha, IL-8, IFNgamma and IL-6 promoters were synthesised from single stranded (ss) oligos (illustrated in Table 1), (TIB Molbiol, Berlin, Germany) and 32P-labeled with 5'gamma-P-ATP (3,000 Ci/mmol, Amersham, Braunschweig, Germany) using the Ready-To-Go-Polynucleotide-Kinase Kit (PNK Kit, Pharmacia LKB, Freiburg, Germany) according to the manufacturer's instructions. To test the binding specificity of the oligos, mutated oligos were used as an additional control. Mutations in the P2 site of the IFNgamma promoter [29] and in the kappa consensus sequence of the TNFalpha promoter [55] have been reported to interfere with sequences, which seem to be crucial for the binding of NFAT and NFkappaB proteins, respectively. Afterwards, the ds oligos were purified via gel filtration using Probe Quant G-50 Micro Columns (Pharmacia) according to the manufacturer's instructions. 
  