FIG. 5.

Nuclear translocation of NF-κB in cisplatin-treated HEI-OC1 auditory cells. A Cytosolic and nuclear fractions from cells treated with 20 μM cisplatin for indicated periods were subjected to 12% SDS polyacrylamide gel electrophoresis and immunoblotted with antibodies specific for NF-κB p65, IκB, VDAC, and β-actin. B Cells were transfected with NF-κB luciferase reporter gene for 24 h, then treated with 20 μM cisplatin for indicated periods. Luciferase activity was measured at indicated time points. *p < 0.05 and ** p < 0.01 by one-way ANOVA, compared with media control group (n = 3). Cells were treated with cisplatin in the presence of 10 ng/ml concentrations of single or combined antibodies specific for TNF-α, IL-1β, and IL-6 for 60 min (C) or 20 ng/ml concentrations of each cytokine alone or in combination for 60 min (D). Cytosolic and nuclear fractions were then immunoblotted with antibodies specific for NF-κB p65, IκB, VDAC, and β-actin.