Fig. 7

β-LAP increased HO-1 and NQO1 expression via upregulation of Nrf2/ARE and PKA/CREB pathways in LPS-stimulated microglia. a Western blot analysis shows the effects of β-LAP on HO-1 and NQO1 protein expression. Cell lysates were obtained from BV2 cells treated with β-LAP (0.5, 1, and 2 μM) with or without LPS (100 ng/ml) for HO-1 or NQO1 (6–16 h). b RT-PCR was performed to determine the HO-1 and NQO1 mRNA expression. Cells were treated with β-LAP (0.5, 1, and 2 μM) for 1 h prior to treatment with LPS (100 ng/ml, for 6 h) and analyzed. Quantification data are shown in the graph (n = 3). c EMSA for Nrf2. Nuclear extracts were prepared from BV2 cells treated with LPS (100 ng/ml, for 3 h) or LPS + β-LAP (1 and 2 μM, pretreatment for 1 h) and incubated with the ARE probe. The arrow indicates a DNA–protein complex of Nrf2. d Effect of β-LAP on ARE-luc reporter gene activity. Cells transfected with the reporter plasmid (ARE-luc) were treated with β-LAP (0.5, 1, and 2 μM) with or without LPS (100 ng/ml) for 6 h, and the reporter gene assay was performed. e Effect of β-LAP on the phosphorylation of CREB. Cell lysates were obtained from BV2 cells treated with β-LAP (0.5, 1, and 2 μM) with or without LPS (100 ng/ml) for 1 h. Quantification data are shown in the graph (n = 3). f EMSA for CREB. Nuclear extracts were prepared from BV2 cells treated with LPS (100 ng/ml, for 3 h) or LPS + β-LAP (1 and 2 μM, pretreatment for 1 h) and incubated with the CRE probe. The bracket indicates a DNA–protein complex of CREB. g Effect of β-LAP on CRE-luc activity. Cells transfected with the reporter plasmid (CRE-luc) were treated with β-LAP (0.5, 1, and 2 μM), with or without LPS (100 ng/ml) for 6 h, and the reporter gene assay was performed. Data are the means ± SEM of three independent experiments. *P < 0.05 vs. control or ^# P < 0.05 vs. the LPS-treated group