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50s were determined by curve fitting with LSW data analysis software (MDL).
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0028168798
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NF-kB gel shift assays were carried out as described (Schindler, U.; Baichwal, V. R. Mol. Cell Biol. 1994, 14, 5820). In summary, IL8/ECV304 cells were stimulated with TNF for 1 h in the absence or presence of compounds and NF-kB. DNA binding activity was determined by gel shift experiments from crude nuclear extracts.
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Human whole blood assay: blood samples in a 96-well microtiter plate were stimulated for 5 h with LPS in the presence or absence of test compounds. Following the incubation period, levels of TNF in the culture supernatants were determined with an ELISA kit (R&D systems).
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