Journal: Journal of Microbiology and Biotechnology

Article Title: Korean Red Pine ( Pinus densiflora ) Bark Extract Attenuates Aβ-Induced Cognitive Impairment by Regulating Cholinergic Dysfunction and Neuroinflammation

doi: 10.4014/jmb.2207.07015

Figure Lengend Snippet: ( A ) Western blot band image, Protein expression levels of ( B ) p-CREB, and ( C ) BDNF. The results shown are mean ± SD ( n = 3). Data were statistically considered at p < 0.05, and different small letters represent statistical difference.

Article Snippet: Brain-derived neurotrophic factor (BDNF) (CSB-PA05775A0Rb) was purchased from Cusabio Biotech (China), and anti-phospho-cAMP response element-binding protein (p-CREB) (9198) and anti-rabbit (7074S) were purchased from Cell Signaling Technology (USA).

Techniques: Western Blot, Expressing

Journal: Neural Regeneration Research

Article Title: Transplantation of human umbilical cord blood mesenchymal stem cells to treat a rat model of traumatic brain injury ★

doi: 10.3969/j.issn.1673-5374.2012.10.004

Figure Lengend Snippet: Neurotrophic factor expression in surrounding injured brain tissues following human umbilical cord blood mesenchymal stem cell transplantation. (A–C) Nerve growth factor (NGF) protein expression (immunohistochemistry, × 200); (D–F) brain-derived neurotrophic factor (BDNF) protein expression (immunohistochemistry, × 200); (G–I) BDNF mRNA expression ( in situ hybridization, × 400). (A, D, G) Results (absorbance) are expressed as mean ± SD from six rats in each group at each time point. a P < 0.05, vs . model group ( t -test was used to specify differences between two groups at the corresponding time points); (B, E, H) model group at 14 days after transplantation; (C, F, I) transplantation group at 14 days after transplantation. Arrows: NGF protein-, BDNF protein-, and BDNF mRNA-positive cells.

Article Snippet: Primary antibodies were incubated overnight at 4°C in wet box with the following dilutions: rat anti-human BrdU monoclonal antibody (1:150; Beijing Boaosen Biotechnology, China), rabbit anti-mouse BDNF (1:150; Boster, Wuhan, China), NGF (1:150; Boster), VEGF (1:150; Boster), CD34 (1:100; Boster), bFGF (1:100; Beijing Boaosen Biotechnology) polyclonal antibody.

Techniques: Expressing, Transplantation Assay, Immunohistochemistry, Derivative Assay, In Situ Hybridization

Journal: BMC Nephrology

Article Title: TL1-A can engage death receptor-3 and activate NF-kappa B in endothelial cells

doi: 10.1186/1471-2369-15-178

Figure Lengend Snippet: Response of vascular endothelial cells in human kidney in organ culture to TL1A. Confocal images of kidney organ culture incubated with either culture media alone or with TL1A (0.2 μg/ml) or TNF (5 ng/ml) for 3 hours at 37°C. (A-C) cultures incubated in media alone show EC staining for NF-κBp65 (green) in both the glomerular (inset) and in peritubular vassels. Some peritubular vessels EC showed positive staining for DR3 (red). (D-F) Cultures treated with TL1A show co-staining for NF-κBp65 (green) and DR3 (red) in EC of some blood vessel (arrows). Inset; show no signal for NF-κBp65 on glomerular EC. (G-I) In contrast, TNF-treated cultures show a strong signal for NF-κB p65 (green) in EC of glomerular (inset) and peritubular blood vessels. DR3 (red) is present only in EC of peritubular blood vessels negative for NF-κB p65. (Original Mags; x40).

Article Snippet: Rabbit anti-human NF-κB p65 was purchased from Serotec (Oxford, U.K).

Techniques: Organ Culture, Incubation, Staining

Journal: Frontiers in Cellular Neuroscience

Article Title: High abundance of BDNF within glutamatergic presynapses of cultured hippocampal neurons

doi: 10.3389/fncel.2014.00107

Figure Lengend Snippet: Two anti-BDNF antibodies enable specific and dense indirect immunolabeling of endogenous BDNF in hippocampal neurons in vitro . (A) Antibody verification with recombinant BDNF. HeLa cells stably expressing lentivirally-delivered BDNF IRES GFP were labeled with different anti (α)-BDNF antibodies from different species (r, rabbit; m, mouse; s, sheep). Antibody details are given in the Material and Method section. As transduction control, GFP was expressed bicistronically under an IRES2 sequence and postlabeled with chicken anti-GFP. BDNF was accumulated in the early secretory trafficking pathway after microtubule disruption with nocodazole, causing a perinuclear Golgi-like staining pattern. (B) Immunoblot of rαBDNF (17 h). Serial dilutions of recombinant BDNF (250–31 pg per lane) served as control (left panel). Hippocampal BDNF harvested at postnatal day 5 was absent in bdnf −/− mice, while heterozygous and wildtype controls show mature BDNF at a M r of 13 kDa. HRP-conjugated anti-Cytochrome C (Cyt C) served as loading control (left panel). In adult mice, rαBDNF (17 h) revealed mature BDNF at high concentration in the hippocampus (left and right lane), while only low amounts were detected in cerebellar protein lysates (middle lane). (right panel). Immunoblots revealed high specificity of the antibody 17 h. (C) Endogenous BDNF in hippocampal neurons at day in vitro (DIV) 35. In somatic regions, BDNF was not concentrated in perinuclear Golgi cisternae, but rather in somatic vesicles. Golgi cisternae were labeled with luminal GFP, a marker for the anterograde, secretory pathway. Luminal GFP and endogenous BDNF co-labeled anterograde trafficking structures (arrow heads) in neurites of hippocampal neurons. Endogenous BDNF was double-labeled by two different anti-BDNF antibodies (rabbit anti-BDNF 17h and mouse anti-BDNF mab9). (D) Somatic anti-BDNF labels were lost when the bdnf gene was removed from single cells. Hippocampal neurons from bdnf fl/fl mice were transduced with a low titer of a lentivirus expressing tdTomato IRES Cre. Neurons (DIV 20) were labeled with anti-BDNF (mab9). Tomato+ cells (upper neuron, red, yellow arrow) lacked a typical vesicular somatic BDNF label, while untransduced, Tomato-Cre-deficient neurons (lower neuron, cyan arrow) exhibited a strong somatic BDNF label. In the composite image Tomato (red) and mouse anti-BDNF (mab9) (green) labels are shown together with an anti-neurofilament (Nfh) label (white) and a nuclear DAPI stain (blue). Bar: 25 μm.

Article Snippet: Blocking and antibody incubation were performed in 10% goat serum, 5% milk powder (Biorad) for 3–4 h. For BDNF detection, rabbit anti-BDNF 17H (1/6000, provided by Michael Sendtner, Institute for Clinical Neurobiology, Würzburg, Germany) was incubated over night at 4°C.

Techniques: Immunolabeling, In Vitro, Recombinant, Stable Transfection, Expressing, Labeling, Transduction, Control, Sequencing, Disruption, Staining, Western Blot, Concentration Assay, Marker

Journal: International Journal of Neuropsychopharmacology

Article Title: Sustained Ultrastructural Changes in Rat Hippocampal Formation After Repeated Electroconvulsive Seizures

doi: 10.1093/ijnp/pyaa021

Figure Lengend Snippet: Brain-derived neurotrophic factor (BDNF) expression levels were measured by immunohistochemistry in hippocampus. (A) BDNF expression in the subregions of hippocampus. (B–D) Immunohistochemistry examined BDNF expression levels in each group. Mean optical density (MOD) was calculated with the following formula: OD = log10(max pixel intensity/mean pixel intensity), where max pixel intensity = 255. A single ECS treatment significantly increased the MOD of BDNF expression in both FSL and FRL rat strains in CA1 of hippocampus in contrast to DG and CA2/3 of hippocampus. No difference between the FRL sham and FSL sham rats was observed. Three months after 10xECS, no changes of BDNF expression levels in hippocampal subregions between the FRL and FSL rats were observed.

Article Snippet: Tissue sections were incubated at 4°C overnight in a solution containing rabbit anti-BDNF polyclonal antibody (1:500) (AB1779, Merck Millipore) or mouse anti-VEGF monoclonal antibody (1:400) (sc-53462, Santa Cruz Biotechnology, Inc.).

Techniques: Derivative Assay, Expressing, Immunohistochemistry

Journal: International Journal of Neuropsychopharmacology

Article Title: Sustained Ultrastructural Changes in Rat Hippocampal Formation After Repeated Electroconvulsive Seizures

doi: 10.1093/ijnp/pyaa021

Figure Lengend Snippet: Results of All Groups in FSL Rats

Article Snippet: Tissue sections were incubated at 4°C overnight in a solution containing rabbit anti-BDNF polyclonal antibody (1:500) (AB1779, Merck Millipore) or mouse anti-VEGF monoclonal antibody (1:400) (sc-53462, Santa Cruz Biotechnology, Inc.).

Techniques:

Journal: International Journal of Neuropsychopharmacology

Article Title: Sustained Ultrastructural Changes in Rat Hippocampal Formation After Repeated Electroconvulsive Seizures

doi: 10.1093/ijnp/pyaa021

Figure Lengend Snippet: Results of All Groups in FRL Rats

Article Snippet: Tissue sections were incubated at 4°C overnight in a solution containing rabbit anti-BDNF polyclonal antibody (1:500) (AB1779, Merck Millipore) or mouse anti-VEGF monoclonal antibody (1:400) (sc-53462, Santa Cruz Biotechnology, Inc.).

Techniques:

Journal: Neurochemistry international

Article Title: DL-3-n-butylphthalide Induced Neuroprotection, Regenerative Repair, Functional Recovery and Psychological Benefits following Traumatic Brain Injury in Mice

doi: 10.1016/j.neuint.2017.03.017

Figure Lengend Snippet: TBI mice received acute as well as daily TBI intranasal treatments until sacrifice. Expressions of regenerative factors were measured using Western blot 21 days after TBI. A. The protein levels of BDNF, VEGF, GDNF, eNOS, CXCR4, and MMP-9. B. Quantified data from A. NBP treatments significantly enhanced the expression of BDNF, VEGF, eNOS, and MMP-9 compared to the control group. There was no significant change in GDNF and CXCR4 expressions among groups. * P<0.05 versus sham group; # P<0.05 versus TBI-saline control group by one-way ANOVA followed by Bonferroni correction; n=5 per group.

Article Snippet: The primary antibodies used and the dilutions for each were rabbit anti-cleaved caspase-3 antibody (Cell Signaling, Danvers, MA) at 1:400, rabbit anti-caspase-9 antibody (Cell Signaling) at 1: 1,000, rabbit anti-cytochrome c antibody (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) at 1:200 or rabbit anti-AIF antibody (Cell Signaling) at 1:500, rabbit anti-TNF-α (Cell Signaling) at 1:1000, rabbit anti-IL-1β (Cell Signaling) at 1:1000, rabbit anti-BDNF (BD Bioscience, San Jose, CA) at 1:2,000, rabbit anti-VEGF (BD Bioscience) at 1,000, rabbit anti-GDNF (BD Bioscience) at 1:1,000, rat anti-eNOS (BD Bioscience) at 1:1,000, rabbit anti-CXCR4 (BD Bioscience) at 1:2,000, rabbit anti-p65 (Cell Signaling) at 1:1,000, mouse anti-IκB (Cell Signaling) at 1:1,000, mouse anti-p-IκB (Cell Signaling) at 1:1,000, rabbit anti-COX (Cell Signaling) at 1:2,500, mouse anti-actin (Sigma-Aldrich) at 1:5000, and rabbit anti-MMP-9 antibody (Millipore, Billerica, MA) at 1:2500.

Techniques: Western Blot, Expressing