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The secondary antibodies were amplified with the Vector Elite ABC kit (1:100; Vector Laboratories Inc.). Antibody-biotin-avidin-peroxidase complexes were visualized using 0.03% diaminobenzidine (DAB), and the sections were mounted onto gelatin-coated slides. The slides were airdried overnight at room temperature, and coverslips were mounted using Permount (Fisher Scientific, Pittsburgh, PA, USA). Western Blot for 5-HT1A Receptors Western blot for the 5-HT1A receptor was performed, according to a previously described method [16,17]. The dorsal raphe was collected and immediately frozen at -70?C. Protein from each dorsal raphe was extracted. The tissues were homogenized with lysis buffer containing 50mM Tris-HCl (pH 8.0), 150mM NaCl, 10% glycerol, 1% Triton X-100, 1.5mM MgCl2��6H2O, [http://www.selleckchem.com/products/BIBW2992.html click here] 1mM ethyleneglycotetraacetic acid, 1mM phenylmethylsulfonyl fluoride, 1mM Na2VO4, and 100mM NaF, then ultracentrifuged at 50,000 rpm for 1 hour. Protein content was measured using a Bio-Rad colorimetric protein assay kit (Bio-Rad, Hercules, CA, USA). Protein (30 ��g) was separated on sodium dodecyl sulfate-polyacrylamide [http://en.wikipedia.org/wiki/SWAP70 SWAP70] gels and transferred onto nitrocellulose membranes. Anti-GAPDH antibody (1:3,000; Abcam) and anti�C5-HT1A antibody (1:1,000; Abcam) were used as the primary antibodies. Horseradish peroxidase-conjugated antimouse antibodies for GAPDH and 5-HT1A were used as the secondary antibodies. Experiments were performed in normal laboratory conditions and at room temperature, except for the transferred membranes. Membrane transfer was performed at 4?C with a cold pack and prechilled buffer. Band detection was performed using the enhanced chemiluminescence detection kit (Amersham Pharmacia Biotech GmbH, Freiburg, Germany). Data Analysis For confirming the expression of 5-HT1A receptors, the detected bands were quantified densitometrically using Molecular Analyst ver. 1.4.1 (Bio-Rad). The numbers of 5-HT and TPH-positive cells in the dorsal raphe were counted hemilaterally under a light microscope (Olympus, Tokyo, Japan). The data were analyzed with one-way analysis of variance followed by the Duncan post hoc test. All values are expressed as the mean��standard error of the mean. P-value [http://www.selleckchem.com/products/BKM-120.html NVP-BKM120 mw] State The results of the forced swimming test are presented in Fig. 1. The average time of immobility in the control, control and exercise, stress-induced, and stress-induced and exercise groups were 9.28��2.00, 4.92��0.98, 78.81��10.67, and 43.21��10.56 seconds, respectively. The average time of climbing in the groups was 189.52��13.00, 180.29��8.90, 20.00��4.07, and 62.08��10.70 seconds, respectively. Induction of stress increased the immobility time and decreased the climbing time (P