Fig. 3

Effect of SA-EE on Nrf2 pathway activation. HT22 cells were exposed to 5 mM glutamate alone or glutamate in combination with 50 μg/mL SA-EE for 1 h prior to western blot analysis of nuclear and cytoplasmic Nrf2 levels (a). Quantitative real-time RT-PCR analysis of NQO1, GCLM and EAAT3 mRNA expression was performed after treatment of cells with 5 mM glutamate alone or glutamate in combination with 50 μg/mL SA-EE or 15 μM curcumin (positive control) for 24 h (b). Whole cell lysates were subjected to western blot analysis of EAAT3 level after treating cells with 5 mM glutamate alone or glutamate in combination with 50 μg/mL SA-EE for 24 h (c). Lamin B1 and β-actin served as endogenous loading controls for nuclear extracts and whole cell/cytoplasmic extracts, respectively. β-actin was used as an internal control for RT-PCR assay. All data were normalized to endogenous control levels and are expressed as the mean ± SEM of at least three independent experiments. ^## P < 0.01, ^### P < 0.001 vs. control; **P < 0.01, ***P < 0.001 vs. glutamate alone