rabbit anti-vgf  (Millipore)

Journal: Neuropsychiatric Disease and Treatment

Article Title: Multisensory Fusion Training and 7, 8-Dihydroxyflavone Improve Amyloid-β-Induced Cognitive Impairment, Anxiety, and Depression-Like Behavior in Mice Through Multiple Mechanisms

doi: 10.2147/NDT.S459891

Figure Lengend Snippet: Increase in BDNF, PSD95, VGF, and p-CREB protein levels in the hippocampus of Aβ-induced mice due to the combined application of DHF and MSFT intervention. ( A ) Western blot expression images of the respective proteins in mouse hippocampal tissue. ( B – J ) Expression levels of BDNF (28 kDa), VGF, PSD-95, Nrf2, p-Ser9-GSK3β, total GSK3α/β, p-CREB and CREB protein in the mouse hippocampal tissue. *P < 0.05, **P < 0.01.

Article Snippet: Subsequently, the PVDF membranes were incubated overnight at a temperature of 4°C with the following antibodies: rabbit anti-BDNF antibody (1:1000, Abcam, ab108319), rabbit anti-VGF antibody (1:1000, Abcam, ab74140), rabbit anti-PSD-95 antibody (1:1000, Cell Signaling Technology, 3450), rabbit anti-Nrf2 antibody (1:1000, Abcam, ab62352), rabbit anti-p-Ser9-GSK3β antibody (1:1000, Cell Signaling Technology, 5558), rabbit anti-GSK3α/β antibody (1:1000, Cell Signaling Technology, 5676), mouse anti-phospho-CREB (Ser133) antibody (1:1000, Cell Signaling Technology, 9196), and anti-beta actin (1:5000, Invitrogen, MA5-15,739).

Techniques: Western Blot, Expressing

Journal: International Journal of Neuropsychopharmacology

Article Title: PI3K/AKT/mTOR Signaling-Mediated Neuropeptide VGF in the Hippocampus of Mice Is Involved in the Rapid Onset Antidepressant-Like Effects of GLYX-13

doi: 10.1093/ijnp/pyu110

Figure Lengend Snippet: Vgf knockdown in hippocampus blocks the rapid-acting antidepressant-like effects of GLYX-13. (a) Experimental procedure for the test schedule. NS shRNA or Vgf shRNA were microinfused into bilateral hippocampus of mice following 7-day acclimatization. GLYX-13 (10mg/kg, i.p.) or its vehicle was administrated beginning from 7 days after the viral infusions (day 1) and then 30 or 60 minutes later, the open field test (OFT) or forced swim test (FST) was conducted, respectively. (b) NS- shRNA or Vgf -shRNA were well expressed in HEK293 cells as indicated by EGFP (green) observed under a fluorescence microscope. Scale bar=100 μm. (c) The results are expressed as 2 -△ Ct [△ Ct = Ct (VGF) – Ct (β-actin)] and normalized by the level of β-actin. (d) Microinjection sites and specific expressions of EGFP (green) in the hippocampus observed under fluorescence microscopy. Scale bars=200 μm. (e) The hippocampus tissues of 2mm in diameter around the injection site were punched out for qRT-PCR. (f) Vgf knockdown in the hippocampus significantly produced the depressive-like behavior and also blocked the antidepressant-like behavior in the FST of mice. (g) All the treatments had no effects on locomotor activity, reflected by the line crossings (left) and rearings (right) in mice. The data are expressed as mean±SEM (n=5 per group for VGF mRNA expression and n=9 per group for behavioral tests). ** P<. 01 (c), compared with NS shRNA group and ** P<. 01 (d), compared with Vehicle+ NS shRNA group; # P<. 05, compared with Vehicle+ NS shRNA group.

Article Snippet: Membranes were then incubated with rabbit anti-phospho-AKT-Ser473 (1:1000; Cell Signaling, Danvers, MA), rabbit anti-total-AKT (1:1000; Cell Signaling), rabbit anti-phospho-mTOR (1:1000; Millipore), rabbit anti-phospho-total-mTOR (1:1000; Abcam, Cambridge, MA), rabbit anti-phospho-eEF2 (1:800; Abcam), rabbit anti-VGF (1:500; Millipore), or anti-β-actin (1:1000; Chemicon) at 4°C overnight.

Techniques: shRNA, Fluorescence, Microscopy, Injection, Quantitative RT-PCR, Produced, Activity Assay, Expressing

Journal: International Journal of Neuropsychopharmacology

Article Title: PI3K/AKT/mTOR Signaling-Mediated Neuropeptide VGF in the Hippocampus of Mice Is Involved in the Rapid Onset Antidepressant-Like Effects of GLYX-13

doi: 10.1093/ijnp/pyu110

Figure Lengend Snippet: PI3K/AKT/mTOR/VGF activity mediates the antidepressant-like effects of GLYX-13 in mice. (a) Experimental procedure for the assessment of the role of PI3K/AKT/mTOR/VGF signaling in the effects of GLYX-13 (10mg/kg, i.p.). Cannula implantations were microinfused into bilateral hippocampus of mice following 7-day acclimatization. Mice were treated with LY294002 (10 nmol/side) and 30 minutes later followed by GLYX-13 (100mg/kg, i.p.) treatment. Then the open field test (OFT) was conducted 30 minutes later and the forced swim test (FST) was conducted 30 minutes after the OFT. (b) Immobility time of mice was measured. Pretreatment with LY294002 reversed the reduction of immobility time produced by GLYX-13. (c-d) All the treatments had no effects on locomotor activity, reflected by the line crossings (c) and rearings (d) in mice. The data are expressed as mean±SEM (n=9 per group). ** P<. 01, compared with ACSF + Vehicle group; ## P<. 01, compared with ACSF + GLYX-13 group.

Article Snippet: Membranes were then incubated with rabbit anti-phospho-AKT-Ser473 (1:1000; Cell Signaling, Danvers, MA), rabbit anti-total-AKT (1:1000; Cell Signaling), rabbit anti-phospho-mTOR (1:1000; Millipore), rabbit anti-phospho-total-mTOR (1:1000; Abcam, Cambridge, MA), rabbit anti-phospho-eEF2 (1:800; Abcam), rabbit anti-VGF (1:500; Millipore), or anti-β-actin (1:1000; Chemicon) at 4°C overnight.

Techniques: Activity Assay, Produced

Journal: International Journal of Neuropsychopharmacology

Article Title: PI3K/AKT/mTOR Signaling-Mediated Neuropeptide VGF in the Hippocampus of Mice Is Involved in the Rapid Onset Antidepressant-Like Effects of GLYX-13

doi: 10.1093/ijnp/pyu110

Figure Lengend Snippet: Effect of GLYX-13 on levels of pAKT, AKT, pmTOR, mTOR, peEF2. and VGF in hippocampus of mice. (a) Representative immunoblots of pAKT, AKT, pmTOR, mTOR, peEF2, and VGF detected by Western blotting with tissues from the hippocampus; the rest of the panels are quantification of the immunoblotting bands of pAKT (b), AKT (c), pmTOR (d), mTOR (e), peEF2 (f), and VGF (g). The data are expressed as mean±SEM (n=3 per group). * P<. 05, ** P<. 01, compared with nonstressed mice treated with vehicle group; # P<. 05, ## P<. 01, compared with stressed mice treated with GLYX-13 group.

Article Snippet: Membranes were then incubated with rabbit anti-phospho-AKT-Ser473 (1:1000; Cell Signaling, Danvers, MA), rabbit anti-total-AKT (1:1000; Cell Signaling), rabbit anti-phospho-mTOR (1:1000; Millipore), rabbit anti-phospho-total-mTOR (1:1000; Abcam, Cambridge, MA), rabbit anti-phospho-eEF2 (1:800; Abcam), rabbit anti-VGF (1:500; Millipore), or anti-β-actin (1:1000; Chemicon) at 4°C overnight.

Techniques: Western Blot

Journal: International Journal of Neuropsychopharmacology

Article Title: PI3K/AKT/mTOR Signaling-Mediated Neuropeptide VGF in the Hippocampus of Mice Is Involved in the Rapid Onset Antidepressant-Like Effects of GLYX-13

doi: 10.1093/ijnp/pyu110

Figure Lengend Snippet: Effects of Vgf -shRNA and/or GLYX-13 (10mg/kg, i.p.) on the level of pAKT, AKT, pmTOR, mTOR, peEF2, and VGF in hippocampus of mice. (a) Representative immunoblots of pAKT, AKT, pmTOR, mTOR, peEF2, and VGF detected by Western blotting with tissues from the hippocampus; the rest of the panels are quantification of the immunoblotting bands of pAKT (b), AKT (c), pmTOR (d), mTOR (e), peEF2 (f), and VGF (g). The data are expressed as mean±SEM (n=3 per group). * P<. 05, ** P<. 01, compared with Vehicle+ NS shRNA treated group; # P<. 05, ## P<. 01, compared with Vehicle+ NS shRNA treated group.

Article Snippet: Membranes were then incubated with rabbit anti-phospho-AKT-Ser473 (1:1000; Cell Signaling, Danvers, MA), rabbit anti-total-AKT (1:1000; Cell Signaling), rabbit anti-phospho-mTOR (1:1000; Millipore), rabbit anti-phospho-total-mTOR (1:1000; Abcam, Cambridge, MA), rabbit anti-phospho-eEF2 (1:800; Abcam), rabbit anti-VGF (1:500; Millipore), or anti-β-actin (1:1000; Chemicon) at 4°C overnight.

Techniques: shRNA, Western Blot

Journal: International Journal of Neuropsychopharmacology

Article Title: PI3K/AKT/mTOR Signaling-Mediated Neuropeptide VGF in the Hippocampus of Mice Is Involved in the Rapid Onset Antidepressant-Like Effects of GLYX-13

doi: 10.1093/ijnp/pyu110

Figure Lengend Snippet: Pretreatment with LY294002 blocks the effects of GLYX-13 on expression of pAKT, AKT, pmTOR, mTOR, peEF2, and VGF in hippocampus of mice. (a) Representative immunoblots of pAKT, AKT, pmTOR, mTOR, peEF2, and VGF detected by Western blotting with tissues from the hippocampus; the rest of the panels are quantifications of the immunoblotting bands of pAKT (b), AKT (c), pmTOR (d), mTOR (e), peEF2 (f), and VGF (g). The data are expressed as mean±SEM (n=3 per group). * P<. 05, ** P<. 01, compared with ACSF + Vehicle group; ## P<. 01, compared with ACSF + GLYX-13 group.

Article Snippet: Membranes were then incubated with rabbit anti-phospho-AKT-Ser473 (1:1000; Cell Signaling, Danvers, MA), rabbit anti-total-AKT (1:1000; Cell Signaling), rabbit anti-phospho-mTOR (1:1000; Millipore), rabbit anti-phospho-total-mTOR (1:1000; Abcam, Cambridge, MA), rabbit anti-phospho-eEF2 (1:800; Abcam), rabbit anti-VGF (1:500; Millipore), or anti-β-actin (1:1000; Chemicon) at 4°C overnight.

Techniques: Expressing, Western Blot