NF-kB activation and function after TNF
treatment of OM-10.1 cells in the presence of benzothiophene derivatives
(a) Nuclear extracts from 30-min TNFα-treated OM-10.1 cultures were interacted with a labeled NF-κB-specific probe either alone (—) or in combination with an 24-fold excess of unlabeled specific (κB) and nonspecific (oct) competitor probes. The bound products of these reactions were identified by electrophoresis and autoradiography. TNFα-induced, NF-κB-specific binding activity is marked by the arrow, (b) Cytoplasmic extracts from OM-10.1 cultures before (T-0) and at various times (30 min and 2 hr) after TNFα treatment were tested by immunoblot analysis for I-κB. A corresponding medium culture (not TNFα treated) was harvested at each time point. The band shown migrated to 37 kD, characteristic of I-κB.