Figure 9.

The effects of steppogenin (1) on IκB-α phosphorylation and degradation (A), NF-κB activation (B,C), and NF-κB localization (D) in LPS-stimulated primary rat microglial cells. (A–D) The cells were pretreated for 3 h with the indicated concentrations of 1 and then stimulated for 1 h with LPS (1 μg/mL). Total proteins were prepared and the western blot analysis was performed using specific IκB-α, p-IκB-α p65, and p50 antibodies. A commercially available NF-κB ELISA (Active Motif) was used to test the nuclear extracts and determine the degree of NF-κB binding. The data are presented as the mean ± SD of three experiments. The band intensities were quantified by densitometry and normalized to the intensity of β-actin or PCNA; the normalized values are presented below each band.