Figure 4

Glyceollin-induced nuclear localization of Nrf2 and HO-1 expression. (A) HT22 cells were incubated with glyceollins (10 μM) and MAPK and PI3K inhibitors for 6 h and separated into nuclear and cytoplasm extracts. Each extract was used for Western blot analysis. Lamin B and β-actin were used as a protein loading control. Inhibitors used are as follows; SP (ERK inhibitor, SP600125), SB (a p38 inhibitor, SB203580), PD (JNK inhibitor, PD98059), or LY (PI3K inhibitor); (B) SH-SY5Y cells were incubated with various concentrations of glyceollins for 12 h, followed by Western blot analysis for HO-1 expression. In addition, SHSY5Y cells were transiently transfected with HO-1 siRNA and subsequently treated with glyceollins for 24 h. Whole cell extract was collected and used for Western blot analysis of HO-1 expression. All experiments were conducted in duplicate.