β-Asarone treatment increases ERK1/2 and CREB phosphorylation in hippocampus of CUMS-exposed rats. Rats were orally treated with β-asarone (25 mg/kg per day) or saline for 28 days while being exposed to CUMS or control conditions. (A) Phospho-protein levels of CREB were determined by immunoblot analysis with antibody against phosphorylated CREB. Total CREB levels showed equivalent protein loading. A representative immunoblot for p-CREB, CREB and GAPDH. (B) The graph demonstrates the densitometric analysis of CREB phosphorylation (normalized to total CREB). Note that the levels of total CREB and GAPDH remained constant. (C) Total RNA was isolated from hippocampus using RNAiso reagent and used for cDNA synthesis. The mRNA levels CREB was detected by real-time PCR and the 2−ΔΔCt method. (D) Phospho-protein levels of ERK1/2 were determined by immunoblot analysis with antibody against phosphorylated ERK1/2. Total ERK levels showed equivalent protein loading. A representative immunoblot for p-ERK1/2, ERK1/2 and GAPDH. (E) The graph demonstrates the densitometric analysis of ERK1/2 phosphorylation (normalized to total ERK1/2). Note that the levels of total ERK1/2 and GAPDH remained constant. Data were presented as mean ± S.D., n = 6 per group. *, p < 0.05 compared with no stress/saline animals; ∫, p < 0.05 compared with CUMS/saline animals (analysis of variance and SNK post hoc test).