Journal: Cell Stem Cell

Article Title: Selective advantage of mutant stem cells in human clonal hematopoiesis is associated with attenuated response to inflammation and aging

doi: 10.1016/j.stem.2024.05.010

Figure Lengend Snippet:

Article Snippet: APC anti-human CD271 (clone ME20.4-1.H4) , Miltenyi , Cat# 130-113-418; RRID: AB_2733363.

Techniques: Recombinant, Reverse Transcription, DNA Library Preparation, DNA Sequencing, RNA Sequencing, Purification, Sequencing, Software, Sterility

Journal: Journal of Clinical Medicine

Article Title: Moderate Aerobic Exercise Regulates Follicular Dysfunction by Initiating Brain-Derived Neurotrophic Factor (BDNF)-Mediated Anti-Apoptotic Signaling Pathways in Polycystic Ovary Syndrome

doi: 10.3390/jcm11195584

Figure Lengend Snippet: Sequences of primers designed for RT-qPCR.

Article Snippet: Sections were incubated overnight at 4 °C with antibodies against BDNF (1:100, Wanleibio), TrkB (1:100, Wanleibio), p75NTR (1:100, 55014-1-AP, Proteintech, Chicago, IL, USA), and Cleaved-Caspase-3 (1:100, 66470-2-Ig, Proteintech) at a 1:100 dilution.

Techniques:

Journal: Journal of Clinical Medicine

Article Title: Moderate Aerobic Exercise Regulates Follicular Dysfunction by Initiating Brain-Derived Neurotrophic Factor (BDNF)-Mediated Anti-Apoptotic Signaling Pathways in Polycystic Ovary Syndrome

doi: 10.3390/jcm11195584

Figure Lengend Snippet: The expression of BDNF, TrkB, and p75NTR in ovarian follicles of PCOS rats after aerobic exercise. Rats received DHEA for the induction of polycystic ovarian syndrome, together with or without exercise treatment. ( A , B ) The expression levels of BDNF in ovarian follicles at different stages were analyzed by immunohistochemistry. ( C , D ) The expression levels of BDNF in ovarian follicles were analyzed by immunofluorescence. ( E , F ) The co-expression of TrkB and p75NTR in ovarian follicles at different stages was detected by immunofluorescence double labeling. n = 10 in each group. Data are shown as mean ± SEM. ** p < 0.05, vs. control group; ## p < 0.01, vs. PCOS group.

Article Snippet: Sections were incubated overnight at 4 °C with antibodies against BDNF (1:100, Wanleibio), TrkB (1:100, Wanleibio), p75NTR (1:100, 55014-1-AP, Proteintech, Chicago, IL, USA), and Cleaved-Caspase-3 (1:100, 66470-2-Ig, Proteintech) at a 1:100 dilution.

Techniques: Expressing, Immunohistochemistry, Immunofluorescence, Labeling, Control

Journal: Journal of Clinical Medicine

Article Title: Moderate Aerobic Exercise Regulates Follicular Dysfunction by Initiating Brain-Derived Neurotrophic Factor (BDNF)-Mediated Anti-Apoptotic Signaling Pathways in Polycystic Ovary Syndrome

doi: 10.3390/jcm11195584

Figure Lengend Snippet: Aerobic exercise activated BDNF signaling in the ovary of DHEA-induced PCOS rats. Rats received DHEA for the induction of polycystic ovarian syndrome, together with or without exercise treatment. ( A ) mRNA expression of BDNF, TrkB, and p75NTR factors in ovarian tissue was analyzed by real-time PCR. ( B , C ) The expression of BDNF, TrkB, and p75NTR in ovarian tissue was assessed by Western blot assay. ( D , E ) The expression of PI3K, AKT, and p-AKT in ovarian tissue was assessed by Western blot assay. ( F – I ) The expression of p75NTR, NF-κB, p-NF-κB, JNK, and p-JNK in ovarian tissue was assessed by Western blot assay. n = 10 in each group. Data are shown as mean ± SEM. ** p < 0.05, vs. control group; ## p < 0.01, vs. PCOS group.

Article Snippet: Sections were incubated overnight at 4 °C with antibodies against BDNF (1:100, Wanleibio), TrkB (1:100, Wanleibio), p75NTR (1:100, 55014-1-AP, Proteintech, Chicago, IL, USA), and Cleaved-Caspase-3 (1:100, 66470-2-Ig, Proteintech) at a 1:100 dilution.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal:

Article Title: Detergent-insoluble GPI-anchored Proteins Are Apically Sorted in Fischer Rat Thyroid Cells, but Interference with Cholesterol or Sphingolipids Differentially Affects Detergent Insolubility and Apical Sorting

doi:

Figure Lengend Snippet: Steady-state distribution and delivery to the plasma membrane of PLAP and NTR-PLAP in transfected FRT cells. (A) Schemes of PLAP and NTR-PLAP (a fusion protein containing the ectodomain of p75NTR and the C-terminal GPI attachment signal of PLAP). (B) FRT cells stably expressing PLAP or NTR-PLAP were grown to confluence and pulsed overnight with [35S]met–cys or [35S]cys, respectively. After surface biotinylation from the apical (Ap) and basolateral (BL) domains, cells were lysed, immunoprecipitated against specific antibodies, and reprecipitated against streptavidin. (C) FRT cells were labeled for 20 min with [35S]met–cys or [35S]cys. At different times of the chase, surface proteins were biotinylated from the apical (Ap) or the basolateral (BL) side. Cells were then lysed and immunoprecipitated with antibodies against PLAP or NTR and with streptavidin beads. Samples were run on 10% SDS-PAGE and detected by fluorography. PLAP and NTR-PLAP are directly sorted to the apical surface in FRT cells, whereas a small percentage of the two proteins is delivered with similar kinetics to the basolateral domain.

Article Snippet: The polyclonal antibody against PLAP was from Rockland (Gilbertsville, PA), and anti-p75NTR mAb was a gift from Andrè Le Bivic (Institut de Biologie du Développement de Marseille, Marseille, France).

Techniques: Transfection, Stable Transfection, Expressing, Immunoprecipitation, Labeling, SDS Page

Journal:

Article Title: Detergent-insoluble GPI-anchored Proteins Are Apically Sorted in Fischer Rat Thyroid Cells, but Interference with Cholesterol or Sphingolipids Differentially Affects Detergent Insolubility and Apical Sorting

doi:

Figure Lengend Snippet: Pulse-chase analysis and purification on sucrose gradients of transmembrane forms of PLAP and p75NTR in transfected FRT cells. (A) Schemes of p75NTR and PLAP-PS321 (a fusion protein containing the ectodomain of PLAP and the transmembrane and the cytoplasmic domain of the PS321 NTR mutant). (B) Pulse-chase and TX-100 extraction of p75NTR and PLAP-PS321. FRT cells expressing p75NTR and PLAP-PS321 were labeled with [35S]cys or [35S]met–cys, respectively, and subsequently treated as described in Figure ​Figure2A.2A. (C) Purification of p75NTR- and PLAP-PS321–enriched fractions on sucrose density gradients. Labeled proteins of FRT cells expressing p75NTR and PLAP-PS321 were solubilized in TX-100 and purified as described in Figure ​Figure2B.2B. Both transmembrane proteins are soluble in TX-100 and do not float on sucrose density gradients.

Article Snippet: The polyclonal antibody against PLAP was from Rockland (Gilbertsville, PA), and anti-p75NTR mAb was a gift from Andrè Le Bivic (Institut de Biologie du Développement de Marseille, Marseille, France).

Techniques: Pulse Chase, Purification, Transfection, Mutagenesis, Expressing, Labeling

Journal:

Article Title: Detergent-insoluble GPI-anchored Proteins Are Apically Sorted in Fischer Rat Thyroid Cells, but Interference with Cholesterol or Sphingolipids Differentially Affects Detergent Insolubility and Apical Sorting

doi:

Figure Lengend Snippet: Surface expression of GPI-anchored, secretory, and transmembrane proteins after FB1 treatment. (A) FRT cells stably expressing PLAP, NTR-PLAP, or gD1-DAF were grown to confluence in the absence (−) or presence (+) of FB1, as described in MATERIALS AND METHODS. After 30 min of labeling with [35S]met–cys or [35S]cys and a 2-h chase, cells were surface biotinylated from the apical (Ap) or the basolateral (BL) domain, lysed, immunoprecipitated against specific antibodies, and reprecipitated against streptavidin. (B) Amounts of labeled proteins were quantified from three independent experiments and are shown as means ± SD. Hatched bars represent apical proteins, and black bars represent basolateral proteins. Apical PLAP and NTR-PLAP, but not basolateral gD1-DAF, became unpolarized after FB1 treatment. (C) FRT cells expressing PLAP-sec, NTR-sec, p75NTR, or PLAP-PS321 were grown to confluence in the absence (−) or presence (+) of FB1, as described in MATERIALS AND METHODS. After labeling with [35S]met–cys or [35S]cys for 20 min and a chase of 2 h, apical and basolateral media were collected separately and cells were surface biotinylated from apical or basolateral domains. Polarized secretion of the secretory forms was revealed by immunoprecipitation in the apical and basolateral medium (top two panels), whereas biotinylated transmembrane proteins were revealed by double immunoprecipitation with specific antibody and streptavidin beads (bottom two panels). (D) Amounts of labeled proteins were quantified from three independent experiments and are shown as means ± SD. Hatched bars represent apical proteins, and black bars represent basolateral proteins. FB1 does not alter the sorting of transmembrane and secretory proteins.

Article Snippet: The polyclonal antibody against PLAP was from Rockland (Gilbertsville, PA), and anti-p75NTR mAb was a gift from Andrè Le Bivic (Institut de Biologie du Développement de Marseille, Marseille, France).

Techniques: Expressing, Stable Transfection, Labeling, Immunoprecipitation

Journal:

Article Title: Detergent-insoluble GPI-anchored Proteins Are Apically Sorted in Fischer Rat Thyroid Cells, but Interference with Cholesterol or Sphingolipids Differentially Affects Detergent Insolubility and Apical Sorting

doi:

Figure Lengend Snippet: Surface expression of GPI-anchored and transmembrane proteins after cholesterol depletion. (A) FRT cells expressing PLAP, NTR-PLAP, gD1-DAF, p75NTR, or PLAP-PS321 were grown on filters and treated with mevinolin (Mev.; 10 μM) for 48 h and M-β-CD (10 mM) for 60 min. Cells were then pulse labeled for 30 min, chased for 2 h, and biotinylated as usual. (B) Amounts of labeled proteins were quantified in three independent experiments and are shown as means ± SD. Hatched bars represent apical (Ap) proteins, and black bars represent basolateral (BL) proteins. Removal of cholesterol does not alter the surface expression of apical and basolateral GPI-anchored and transmembrane proteins.

Article Snippet: The polyclonal antibody against PLAP was from Rockland (Gilbertsville, PA), and anti-p75NTR mAb was a gift from Andrè Le Bivic (Institut de Biologie du Développement de Marseille, Marseille, France).

Techniques: Expressing, Labeling

Journal: FEBS Open Bio

Article Title: The p75 NTR and its carboxyl‐terminal fragment exert opposing effects on melanoma cell proliferation and apoptosis via modulation of the NF‐κB pathway

doi: 10.1002/2211-5463.13047

Figure Lengend Snippet: The effects of p75 NTR ‐CTF on cell proliferation and sorafenib resistance. (A) A375 cells were transfected with pCMV‐p75 NTR ‐FL and pCMV‐p75 NTR ‐CTF. Immunoblot results of endogenous and exogenous expressions of p75 NTR ‐FL (75 kDa) and p75 NTR ‐CTF (25 kDa) in A375 cells. (B) Relative mRNA expression levels of cell‐cycle‐related genes ( CDK2 , CDK4 , cyclin D1 , cyclin B1 and cdc2 ) were assessed by qRT‐PCR. (C) A375 cells were treated with sorafenib of different doses (0, 2, 4, 8 and 16 µ m ) for 24, 48 or 72 h. The effects of sorafenib on A375 cell survival were determined by CCK‐8 assay. (D–F) Stably overexpressed A375 cell lines of p75 NTR ‐FL, p75 NTR ‐CTF and vector were treated with 8 µ m sorafenib. Cell cycle was determined by flow cytometry analysis (D), and cell proliferation was determined by CCK‐8 assay (E) and crystal violet staining (F). All values are presented as the mean ± SD from three independent research results; comparison was performed with Student's t ‐test. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: p75 NTR ‐FL (aa 1–427) and p75 NTR ‐CTF (aa 231–427) sequences of p75 NTR were amplified by PCR from a human cDNA library and cloned into vector pLJM1 (Addgene #19319, Watertown, MA, USA) for expression.

Techniques: Transfection, Western Blot, Expressing, Quantitative RT-PCR, CCK-8 Assay, Stable Transfection, Plasmid Preparation, Flow Cytometry, Staining, Comparison

Journal: FEBS Open Bio

Article Title: The p75 NTR and its carboxyl‐terminal fragment exert opposing effects on melanoma cell proliferation and apoptosis via modulation of the NF‐κB pathway

doi: 10.1002/2211-5463.13047

Figure Lengend Snippet: The effects of p75 NTR ‐CTF on sorafenib‐induced cell apoptosis in vivo and in vitro . (A) Apoptosis was evaluated by flow cytometry after transfection and treatment with sorafenib as indicated. (B) Immunoblot analysis of expression of apoptosis‐associated proteins (caspase‐9, Ccspase‐3, Bax and Bcl‐2) after sorafenib treatment. (C) Nude mouse tumorigenesis test was used to determine cell proliferation and sorafenib resistance of stable expression p75 NTR ‐FL and p75 NTR ‐CTF cell lines. All values shown are mean ± SD of triplicate measurements; comparison was performed with Student's t ‐test. ** P < 0.01; *** P < 0.001.

Article Snippet: p75 NTR ‐FL (aa 1–427) and p75 NTR ‐CTF (aa 231–427) sequences of p75 NTR were amplified by PCR from a human cDNA library and cloned into vector pLJM1 (Addgene #19319, Watertown, MA, USA) for expression.

Techniques: In Vivo, In Vitro, Flow Cytometry, Transfection, Western Blot, Expressing, Comparison

Journal: FEBS Open Bio

Article Title: The p75 NTR and its carboxyl‐terminal fragment exert opposing effects on melanoma cell proliferation and apoptosis via modulation of the NF‐κB pathway

doi: 10.1002/2211-5463.13047

Figure Lengend Snippet: Activation of NF‐κB signaling pathway by p75 NTR ‐CTF. (A) Subcellular localization of NF‐κB p65 (red) was detected by immunofluorescence. Nuclei were stained with DAPI (blue). Scale bar: 100 μm. (B) Cell lysates were extracted from cytosolic and nuclear fractions; the expression levels of p65, β‐actin (cytosolic control) and Lamin B1 (nuclear control) were analyzed by immunoblot analysis. (C) Phosphorylation and total levels of IκBα were assessed by immunoblot analysis. (D) mRNA levels of the antiapoptosis ( c‐IAP1/2 , FLIP ) and tumor angiogenesis ( IL8 , VEGF and bFGF ) were detected by qRT‐PCR. (E) Protein levels of c‐IAP2 and bFGF were measured by western blot analysis. (F, G) Protein levels of IL‐8 and VEGF in the culture media of the cells were measured by ELISA. Data represent the mean ± SD of three separate experiments. Differences between two groups were compared using Student's t ‐test. * P < 0.05; ** P < 0.01; *** P < 0.001. Scale bar: 100 μm.

Article Snippet: p75 NTR ‐FL (aa 1–427) and p75 NTR ‐CTF (aa 231–427) sequences of p75 NTR were amplified by PCR from a human cDNA library and cloned into vector pLJM1 (Addgene #19319, Watertown, MA, USA) for expression.

Techniques: Activation Assay, Immunofluorescence, Staining, Expressing, Control, Western Blot, Phospho-proteomics, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Experimental and Therapeutic Medicine

Article Title: Sevoflurane-induced memory impairment in the postnatal developing mouse brain

doi: 10.3892/etm.2018.5950

Figure Lengend Snippet: Levels of BDNF, Pro-BDNF, TrkB, Akt/PKB and p75NTR in the hippocampal tissues in each group.

Article Snippet: The levels of BDNF, Ntrk2, pro-BDNF, p75NTR and PKB/Akt were measured using an ELISA kit (cat. no. EK0312; Wuhan Boster Bio-Engineering Co., Ltd., Wuhan, China) according to the manufacturer's instructions.

Techniques: Control

Journal: ASN NEURO

Article Title: Inflammatory cytokines IL-1β and TNF-α regulate p75 NTR expression in CNS neurons and astrocytes by distinct cell-type-specific signalling mechanisms

doi: 10.1042/AN20090009

Figure Lengend Snippet: ( A – D ) Cultured hippocampal neurons were treated with either ( A ) IL-1β (10 ng/ml) or ( B ) TNF-α (10 ng/ml) for 4, 8, 12 or 24 h, and were then lysed and analysed by Western blot for p75 NTR and actin. ( C ) and ( D ) Quantification of blots from three experiments as in ( A ) and ( B ) respectively. Densitometric values were normalized to actin and are expressed relative to the untreated cells (time 0). ( E – H ) Cultured hippocampal astrocytes were treated with either ( E ) IL-1β (10 ng/ml) or ( F ) TNF-α (10 ng/ml) for 1, 4, 8, 12 or 24 h, and were then lysed and analysed by Western blot for p75 NTR and actin. ( G ) and ( H ) Quantification of blots from three experiments as in ( E ) and ( F ) respectively. Densitometric values were normalized to actin and are expressed relative to the untreated cells (time 0). ( I and J ) Quantitative real-time PCR analysis of p75 NTR mRNA in hippocampal neurons ( I ) or astrocytes ( J ) treated with IL-1β or TNF-α expressed relative to untreated control cultures. The significance was determined by ANOVA with Tukey's post-hoc analysis. * indicates values significantly different from control at P <0.05.

Article Snippet: Membranes were blocked in 5% (w/v) non-fat dried skimmed milk in TBST [TBS (Tris-buffered saline; 10 mM Tris and 150 mM NaCI, pH 7.4) containing 0.05% Tween 20] and then probed with antibodies against p75 NTR (Upstate Biotechnology), actin (Sigma), p-p38 [phosphorylated-p38 MAPK (mitogen-activated protein kinase)], p-IκB {phosphorylated-IκB [inhibitor of NF-κB (nuclear factor κB)]; Cell Signaling Technology}. p-p38 and p-IκB blots were stripped and reprobed with anti-p38 and anti-IκB (Cell Signaling Technology) antibodies respectively.

Techniques: Cell Culture, Western Blot, Real-time Polymerase Chain Reaction, Control

Journal: ASN NEURO

Article Title: Inflammatory cytokines IL-1β and TNF-α regulate p75 NTR expression in CNS neurons and astrocytes by distinct cell-type-specific signalling mechanisms

doi: 10.1042/AN20090009

Figure Lengend Snippet: ( A ) Hippocampal neurons were pretreated with SB203580 (10 μM) or SN-50 (10 μM) for 30 min and then 10 ng/ml IL-1β was added to the culture for 8 h. Western blot analysis was used to assess p75 NTR and actin levels. ( B ) Quantification of three blots as shown in ( A ). * indicates values are significantly different from control, P <0.05; ** indicates IL-1β+SB203580 is significantly different from IL-1 alone or IL-1β+SN-50, P <0.05. ( C and D ) IL-1 induces p75 NTR through both the NF-κB and p38 MAPK pathways in astrocytes. Cultured astrocytes were pretreated for 30 min with the p38 MAPK inhibitor SB203580 (10 μM) or the NF-κB inhibitor SN-50 (10 μM) prior to IL-1β treatment for 8 h. Western blot analysis was used to detect the p75 NTR and actin levels. ( D ) Quantification of three blots as shown in ( C ). * indicates values are significantly different from control, P <0.05; ** indicates IL-1β+SB203580 and IL-1β+SN-50 are significantly different from IL-1 alone, P <0.05. C, control; SB, SB203580.

Article Snippet: Membranes were blocked in 5% (w/v) non-fat dried skimmed milk in TBST [TBS (Tris-buffered saline; 10 mM Tris and 150 mM NaCI, pH 7.4) containing 0.05% Tween 20] and then probed with antibodies against p75 NTR (Upstate Biotechnology), actin (Sigma), p-p38 [phosphorylated-p38 MAPK (mitogen-activated protein kinase)], p-IκB {phosphorylated-IκB [inhibitor of NF-κB (nuclear factor κB)]; Cell Signaling Technology}. p-p38 and p-IκB blots were stripped and reprobed with anti-p38 and anti-IκB (Cell Signaling Technology) antibodies respectively.

Techniques: Western Blot, Control, Cell Culture

Journal: ASN NEURO

Article Title: Inflammatory cytokines IL-1β and TNF-α regulate p75 NTR expression in CNS neurons and astrocytes by distinct cell-type-specific signalling mechanisms

doi: 10.1042/AN20090009

Figure Lengend Snippet: ( A ) Hippocampal neurons were pretreated with SB203580 (10 μM) or SN-50 (10 μM) for 30 min and then 10 ng/ml TNF-α was provided for 8 h. Western blot analysis was used to assess p75 NTR and actin levels. ( B ) Quantification of three blots as shown in ( A ). * indicates levels are significantly different from control, P <0.05; ** indicates TNFα+SN-50 is significantly different from TNF-α alone, P <0.05. ( C ) Hippocampal astrocytes were pretreated with SB203580 (10 μM) or SN-50 (10 μM) for 30 min and then 10 ng/ml TNFα was added to the culture for 8 h. Western blot analysis was used to assess p75 NTR and actin. ( D ) Quantification of three blots as shown in ( C ). * indicates levels are significantly different from control, P <0.05; ** indicates TNFα+SN-50 is significantly different from TNF-α alone, P <0.05. SB, SB203580. C, control.

Article Snippet: Membranes were blocked in 5% (w/v) non-fat dried skimmed milk in TBST [TBS (Tris-buffered saline; 10 mM Tris and 150 mM NaCI, pH 7.4) containing 0.05% Tween 20] and then probed with antibodies against p75 NTR (Upstate Biotechnology), actin (Sigma), p-p38 [phosphorylated-p38 MAPK (mitogen-activated protein kinase)], p-IκB {phosphorylated-IκB [inhibitor of NF-κB (nuclear factor κB)]; Cell Signaling Technology}. p-p38 and p-IκB blots were stripped and reprobed with anti-p38 and anti-IκB (Cell Signaling Technology) antibodies respectively.

Techniques: Western Blot, Control

Journal: ASN NEURO

Article Title: Inflammatory cytokines IL-1β and TNF-α regulate p75 NTR expression in CNS neurons and astrocytes by distinct cell-type-specific signalling mechanisms

doi: 10.1042/AN20090009

Figure Lengend Snippet: Cultured hippocampal neurons ( A ) or astrocytes ( B ) were treated with IL-1β or TNF-α for 8 h, and incubated with biotin for 1 h. Cell lysates were precipitated with streptavidin. Biotinylated cell-surface protein and non-biotinylated intracellular proteins were analysed by Western blotting for p75 NTR . Blots were stripped and re-probed for actin, which was only present in the intracellular fraction. CTRL, control.

Article Snippet: Membranes were blocked in 5% (w/v) non-fat dried skimmed milk in TBST [TBS (Tris-buffered saline; 10 mM Tris and 150 mM NaCI, pH 7.4) containing 0.05% Tween 20] and then probed with antibodies against p75 NTR (Upstate Biotechnology), actin (Sigma), p-p38 [phosphorylated-p38 MAPK (mitogen-activated protein kinase)], p-IκB {phosphorylated-IκB [inhibitor of NF-κB (nuclear factor κB)]; Cell Signaling Technology}. p-p38 and p-IκB blots were stripped and reprobed with anti-p38 and anti-IκB (Cell Signaling Technology) antibodies respectively.

Techniques: Cell Culture, Incubation, Western Blot, Control

Journal: ASN NEURO

Article Title: Inflammatory cytokines IL-1β and TNF-α regulate p75 NTR expression in CNS neurons and astrocytes by distinct cell-type-specific signalling mechanisms

doi: 10.1042/AN20090009

Figure Lengend Snippet: Schematic diagram showing the signalling pathways activated by IL-1β and TNF-α in neurons (red) and astrocytes (black) leading to induction of p75 NTR .

Article Snippet: Membranes were blocked in 5% (w/v) non-fat dried skimmed milk in TBST [TBS (Tris-buffered saline; 10 mM Tris and 150 mM NaCI, pH 7.4) containing 0.05% Tween 20] and then probed with antibodies against p75 NTR (Upstate Biotechnology), actin (Sigma), p-p38 [phosphorylated-p38 MAPK (mitogen-activated protein kinase)], p-IκB {phosphorylated-IκB [inhibitor of NF-κB (nuclear factor κB)]; Cell Signaling Technology}. p-p38 and p-IκB blots were stripped and reprobed with anti-p38 and anti-IκB (Cell Signaling Technology) antibodies respectively.

Techniques:

Journal: Neural Regeneration Research

Article Title: Association between p75 neurotrophin receptor gene expression and cell apoptosis in tissues surrounding hematomas in rat models of intracerebral hemorrhage ★

doi: 10.3969/j.issn.1673-5374.2012.08.005

Figure Lengend Snippet: Hematoxylin-eosin (HE) and immunohistochemical staining of brain tissue surrounding hematomas after intracerebral hemorrhage (ICH). (A) The yellow arrow indicates neuronal apoptosis as shown by darkly stained cytoplasm and chromatin margination around the hematoma. The blue arrow shows the infiltration of neutrophils, suggesting that inflammation occurs around the hematoma after ICH (HE staining, × 400). (B) After ICH, brain tissue edema around the hematoma occurs as indicated by light staining and cell numbers decrease close to the hematoma, (HE staining, × 200). (C) Perivascular edema, suggesting vascular edema after ICH (HE staining, × 400). (D) p75 neurotrophin receptor is mainly expressed in the cytoplasm of neurons in the control (HE staining, × 400). (E–H) Immunohistochemical staining at 6, 24, 72 hours, and 10 days after ICH, respectively (× 400).

Article Snippet: Immunohistochemical procedures were performed according to the protocol for the p75 NTR immunohistochemistry kit (Promega, Madison, WI, USA).

Techniques: Immunohistochemical staining, Staining, Control

Journal: Foods

Article Title: Increased Levels of Phosphorylated-P38α Induce WNT/β-Catenin and NGF/P75NTR/TrkA Pathways Disruption and SN56 Cell Death following Single and Repeated Chlorpyrifos Treatment

doi: 10.3390/foods13152427

Figure Lengend Snippet: Analysis of CPF action on p-P38α ( A ), P75 NTR ( B ), and PAI-1 ( C ) levels. Results are shown as a percentage of control values. Results of protein content were normalized by total protein concentrations. *** p ≤ 0.001 compared to control. &&& p ≤ 0.001 compared to P38α -silenced cells exposed to CPF.

Article Snippet: To test this hypothesis, P75 NTR - and P38α -silenced or wild-type BF cholinergic SN56 cells were treated singly and repeatedly with CPF (0.1 μM to 50 μM; Sigma, Madrid, Spain) either alone or in combination with recombinant NGF (rNGF; MyBioSource, San Diego, CA, USA) and β-Catenin (rβ-Catenin; MyBioSource, San Diego, CA, USA) proteins.

Techniques: Control

Journal: Foods

Article Title: Increased Levels of Phosphorylated-P38α Induce WNT/β-Catenin and NGF/P75NTR/TrkA Pathways Disruption and SN56 Cell Death following Single and Repeated Chlorpyrifos Treatment

doi: 10.3390/foods13152427

Figure Lengend Snippet: Results from cell viability (MTT assay) ( A ) and apoptosis (caspase 3/7 activity assay) ( B ). Results were normalized by total protein concentrations. Results are shown as a percentage of control values. *** p ≤ 0.001 compared to control. ### p ≤ 0.001 compared to CPF treatment. &&& p ≤ 0.001 compared to P75 NTR -silenced cells treated with CPF. γγγ p ≤ 0.001 compared to rβ-Catenin and CPF treatment. τττ p ≤ 0.001 compared to P38α -silenced cells treated with CPF.

Article Snippet: To test this hypothesis, P75 NTR - and P38α -silenced or wild-type BF cholinergic SN56 cells were treated singly and repeatedly with CPF (0.1 μM to 50 μM; Sigma, Madrid, Spain) either alone or in combination with recombinant NGF (rNGF; MyBioSource, San Diego, CA, USA) and β-Catenin (rβ-Catenin; MyBioSource, San Diego, CA, USA) proteins.

Techniques: MTT Assay, Activity Assay, Control

Journal: The Journal of Neuroscience

Article Title: p75 Neurotrophin Receptor Protects Primary Cultures of Human Neurons against Extracellular Amyloid β Peptide Cytotoxicity

doi: 10.1523/JNEUROSCI.23-19-07385.2003

Figure Lengend Snippet: Anti-extracellular p75 antibody specifically induces neuronal cell death in the presence of Aβ. A, TUNEL-positive cell death in vector-, p75S-, or p75AS-microinjected neurons treated for 24 hr with anti-p75ec. B, TUNEL-positive cell death in serum-deprived neurons treated with anti-p75ec or with anti-p75ic in the absence [control (Ctl)] or presence of 100 nm extracellular Aβ1-42. The difference between Aβ1-42 alone versus p75 antibodies plus Aβ1-42 was assessed by one-way ANOVA followed by Scheffé's test; *p < 0.003. The increase in cell death with anti-p75ic is not significantly different from control. C, Competition of anti-p75ec-induced Aβ toxicity by R-p75NTR; *p < 0.02 compared with Ctl (one-way ANOVA). D, TUNEL-positive cell death in neurons treated for 24 hr with 100 nm Aβ1-42, 0.1 μm H2O2, or 20 μm etoposide in the absence or presence of 1 μg/ml anti-p75ec. Data represent mean and SD.

Article Snippet: An antibody directed against the intracellular segment of p75 NTR (anti-p75ic) (Promega) was used at 1 μg/ml as a control.

Techniques: TUNEL Assay, Plasmid Preparation, Control

Journal: The Journal of Neuroscience

Article Title: p75 Neurotrophin Receptor Protects Primary Cultures of Human Neurons against Extracellular Amyloid β Peptide Cytotoxicity

doi: 10.1523/JNEUROSCI.23-19-07385.2003

Figure Lengend Snippet: p75NTR does not protect human neurons through Akt activation. A, Western blot analyses with phospho-Akt (pAkt) 473, total Akt, or β-actin in neurons treated with 10 μm wortmannin for 6 hr. B, TUNEL-positive cell death in neurons pretreated 1 hr with 10 μm wortmannin and microinjected with wild-type Akt (Akt WT), constitutively active Akt (Akt active), or dominant-negative Akt (Akt DN) before a 24 hr treatment with extracellular Aβ1-42 and wortmannin. Controls are serum deprived [-serum (-S)] for 96 hr after microinjection with the Akt constructs. Data represent the mean and SEM of three independent experiments. C, Western blot analyses of phospho-Akt (P-Akt) 473 or 308, total Akt, and β-actin in proteins from neurons treated with p75ic or p75ec antibodies in the absence or presence of 100 nm Aβ1-42. D, Western blot of phospho-Akt (P-Akt) 308 in proteins from neurons treated with anti-p75ec in the presence of 200 nm or 10 μm wortmannin. Wtm, Wortmannin; Ctl, control.

Article Snippet: An antibody directed against the intracellular segment of p75 NTR (anti-p75ic) (Promega) was used at 1 μg/ml as a control.

Techniques: Activation Assay, Western Blot, TUNEL Assay, Dominant Negative Mutation, Microinjection, Construct, Control