Figure 8.

Time-course effect of nepalolide A on the electrophoretic mobility shift assay (EMSA) for NF-κB binding. C6 glioma cells (a) and astrocytes (b) were incubated with 5 ng ml⁻¹ TNF-α/5 ng ml⁻¹ IL-1β/100 units ml⁻¹ IFN-γ in the presence or absence of 10 μM nepalolide A for 0–180 min. After these times, nuclear extracts were isolated and EMSA for NF-κB binding was performed by using 5′-end-labelled consensus oligonucleotide. The top parts of (a and b) are representative autoradiographs of NF-κB binding. The bottom parts are the relative levels of NF-κB binding in untreated cells or cells treated with nepalolide A. Results are means±s.d. (where large enough to be shown) from four independent experiments, and expressed relative to the control cells treated with mixed cytokines for 3 h. Significant differences between control and nepalolide A-treated cells are indicated by ^*P<0.05; ^**P<0.01; and ^***P<0.001.