Hippocampal and Prefrontal Protein Analysis

Western blotting of hippocampal and prefrontal cortex proteins was performed as described previously 12 , 19 (link). The antibodies used in the experiments were included as following, Goat anti-mouse IgG-HRP (#ZDR-5307, dilution 1:5000) and goat anti-rabbit IgG-HRP (#ZDR-5306, dilution 1:5000) antibodies were purchased from ZSGB-BIO. Anti-Akt (#9272, dilution 1:1000), p-Akt (Ser473) (#4060, dilution 1:1000), Bcl-2 (#3498, dilution 1:5000), BDNF (#47808, dilution 1:1000), TrkB (#4606, dilution 1:1000), p-TrkB (#4619, dilution 1:1000), Cleaved- Caspase3 (#9661, dilution 1:1000) and β-actin (#4970, dilution 1:3000) antibodies were provided by Cell Signaling Technology, Inc. Anti fibroblast growth factor 2 (FGF2) (#sc-136255, dilution 1:1000) antibody was provided by Santa Cruz Biotechnology, Inc. The expression of the target protein was detected and photographed in the Odyssey CLx infrared fluorescence imaging system (LI-COR Biosciences). The relative protein expression levels are quantified by the density-value of the bands and normalized to loading control.