Journal: Journal of diabetes, metabolic disorders & control

Article Title: Deletion of the Neurotrophin Receptor p75 NTR Prevents Diabetes-Induced Retinal Acellular Capillaries in Streptozotocin-Induced Mouse Diabetic Model

doi: 10.15406/jdmdc.2017.04.00129

Figure Lengend Snippet: A summary of sources of antibodies used to detect protein expression using Western Blot.

Article Snippet: Band intensity was quantified using densitometry software (alphEaseFC) and normalized to β-actin. table ft1 table-wrap mode="anchored" t5 caption a7 Antibodies Catalog Number Company Anti-VEGF Cat.# 676472 Millipore, Billerica, MA Anti-EFNB2 Cat #: PA5-19361 Anti-proNGF Cat.# N-250 Alomone Labs, Jerusalem, Israel Anti-NGF Cat.# AN-240 Anti-IL-1β Cat.# ab9722 Abcam, Cambridge, MA Anti-GAPDH Cat.# G8795 Sigma-Aldrich, St. Louis, MO Anti-b-actin Cat.# A2228 Anti-JNK Cat.# sc-7345 Santa Cruz Biotechnology, Dallas, Texas Anti-p-JNK Cat.# sc-293136 Cleaved-PARP Cat.# 5625 Cell Signaling Technology, Danvers, MA Anti-p75 NTR ## Dr. Bruce Carter (Dept. of Biochemistry, Vanderbilt University Open in a separate window GAPDH: Glyceraldehyde 3-phosphate Dehydrogenase; IL-1 β: Interleukin-1 beta; JNKC: June Kinase; NGF: Nerve Growth Factor; proNGF: Proform of Nerve Growth Factor; p75 NTR : Neurotrophin Receptor p75; TrkA: Tyrosine Kinase Receptor-A; VEGF: Vascular Endothelial Growth Factor A summary of sources of antibodies used to detect protein expression using Western Blot.

Techniques: Expressing, Western Blot

Journal: Alzheimer's & Dementia

Article Title: Nerve growth factor (NGF) pathway biomarkers in Down syndrome prior to and after the onset of clinical Alzheimer's disease: A paired CSF and plasma study

doi: 10.1002/alz.12229

Figure Lengend Snippet: Upregulation of proNGF (nerve growth factor) in plasma and cerebrospinal fluid (CSF) from people with Down syndrome (DS). A, The 27 kDa form of proNGF, measured by Western blotting, is upregulated in plasma from Alzheimer's disease (AD)‐asymptomatic people with Down syndrome (aDS) and in Down syndrome with symptomatic AD (DSAD) compared to non‐trisomic controls, one‐way analysis of variance (ANOVA), F = 5.03, P = .01 and Bonferroni post hoc correction; * P < .05. B, ProNGF (27 kDa) is also upregulated in CSF from people with aDS and DSAD compared to non‐trisomic controls, Kruskal Wallis test P = .0002 H = 16.83 and Dunn's post hoc correction; ** P < .01; *** P < .001. C, The 50 kDa form of proNGF is upregulated in plasma from people with aDS and DSAD compared to non‐trisomic controls, one‐way ANOVA, F = 5.62, P = .006 and Bonferroni post hoc correction; * P < .05. D, 50 kDa proNGF is upregulated in the CSF of people with aDS compared to non‐trisomic controls, and further upregulated in people with DSAD, one‐way ANOVA, F = 21.17, P < .0001 and Bonferroni post‐hoc correction; *** P < .001. Data are displayed in box and whisker plots, in which the median is represented by the horizontal line and the whiskers go from each quartile to the minimum or maximum value. HC, Healthy controls: n = 16, aDS: n = 14, DSAD: n = 22; IOD, integrated optical density

Article Snippet: Quantifiability across physiologically relevant concentrations/integrated optical density (IOD) values was established by reconstitution experiments in which plasma/CSF was spiked with 2, 5, 10, or 20 ng of human recombinant proNGF protein (Alomone Labs, Israel), and subsequently analyzed by Western blotting as described above.

Techniques: Western Blot, Whisker Assay

Journal: Alzheimer's & Dementia

Article Title: Nerve growth factor (NGF) pathway biomarkers in Down syndrome prior to and after the onset of clinical Alzheimer's disease: A paired CSF and plasma study

doi: 10.1002/alz.12229

Figure Lengend Snippet: Cerebrospinal fluid (CSF) matrix metalloproteinase (MMP)‐9 and proNGF (precursor of nerve growth factor) discriminate effectively between Alzheimer's disease (AD) symptomatic and asymptomatic people with Down syndrome (DS). A, CSF levels of the 50 kDa proNGF form effectively identify people with Down syndrome with symptomatic Alzheimer's disease (DSAD) from within the total DS sample group, with an area under the receiver operator characteristic (ROC) curve of 0.86. B, CSF MMP‐9 levels effectively identify people with DSAD from within the total DS sample group, with an area under the ROC curve of 0.87. C, ROC curves for the core CSF biomarkers of AD demonstrate that MMP‐9 and proNGF (50 kDa) measured in CSF outperformed CSF pTau‐181 (AUC = 0.85) and total tau (AUC = 0.75), while being outperformed by the Aβ42/40 ratio (AUC = 0.88)

Article Snippet: Quantifiability across physiologically relevant concentrations/integrated optical density (IOD) values was established by reconstitution experiments in which plasma/CSF was spiked with 2, 5, 10, or 20 ng of human recombinant proNGF protein (Alomone Labs, Israel), and subsequently analyzed by Western blotting as described above.

Techniques:

Journal: eNeuro

Article Title: Cortical Brain Injury Causes Retrograde Degeneration of Afferent Basal Forebrain Cholinergic Neurons via the p75NTR

doi: 10.1523/ENEURO.0067-23.2023

Figure Lengend Snippet: Proneurotrophins are induced in the ipsilateral cortex but not the basal forebrain after cortical FPI. a–c , Brain tissue lysates from naive, sham, and injured (2 atm) wild-type adult mice were obtained 1DPI, 3DPI, and 7DPI to determine levels of proBDNF ( a , b ) and proNGF ( c ) in the injured versus uninjured side. Cortical tissue lysate ( a ) harvested for Western blot was probed for proBDNF (32 kDa) in the ipsilateral and contralateral cortex at 1DPI, 3DPI, and 7DPI in naive, sham, and injured mice. Basal forebrain tissue lysate ( b ) harvested for Western blot was probed for proBDNF (32 kDa) in the ipsilateral versus contralateral basal forebrain at 1DPI, 3DPI, and 7DPI in naive, sham, and injured mice. Cortex and basal forebrain tissue lysates ( c ) harvested for Western blot were probed for proNGF (37 kDa) at 3DPI after FPI in the ipsilateral versus contralateral side of the cortex and the basal forebrain; n = 4 (naive), n = 4 (sham 1DPI), n = 4 (injured 1DPI), n = 4 (sham 3DPI), n = 4 (injured 3DPI), n = 3 (sham 7DPI), n = 3 (injured 7DPI; a , b ); n = 3 (naive), n = 4 (sham 3DPI), n = 4 (injured 3DPI; c ). The established size of proBDNF is 32 kDa; however, a prominent band of 25 kDa was also recognized by the BDNF antibody that appeared to be regulated by injury, but the identity of that band is unclear.

Article Snippet: Recombinant human proNGF (cleavage resistant) protein (catalog #N-285) and recombinant mouse proBDNF (cleavage resistant) protein (catalog #B-243) were purchased from Alomone Labs. Poly- d -lysine, glucose, transferrin, insulin, putrescine, selenium, progesterone, penicillin, and streptomycin were purchased from Sigma-Aldrich.

Techniques: Western Blot