Fig. 5.

Glaucine inhibits the NF-κB activity in PMA-stimulated MCF-7 cells. a MCF-7 cells were pretreated with the indicated concentration of glaucine for 30 min and incubated with 100 nM PMA for 30 min. Nuclear extracts were prepared and incubated with radiolabeled oligonucleotides containing the NF-κB motif in the MMP-9 promoter. b MCF-7 cells were pretreated with the indicated concentration of glaucine for 30 min and incubated with 100-nM PMA for 30 min. Cells were harvested and fractionated into the cytoplasm and the nucleus. Lysates were then separated on a 10 % SDS–polyacrylamide gel and subjected to western blotting with anti-p65, anti-p-IκBα, and anti-IκBα antibodies. The analysis was repeated in three times, and α-tubulin and PCNA were used as markers for the cytoplasmic and nuclear fractions. The NF-κB protein level that was translocated to the nucleus was quantified by densitometric analyses. The bars represent the mean ± SD. One-way ANOVA was performed to determine statistical significance (^* P < 0.05)