rabbit polyclonal anti human p75 ntr  (Alomone Labs)

Journal: Cells

Article Title: Etiological Roles of p75 NTR in a Mouse Model of Wet Age-Related Macular Degeneration

doi: 10.3390/cells12020297

Figure Lengend Snippet: MPCs expressing p75 NTR migrate into the laser-injured area of RPE-Choroid. p75 NTR co-localizes in F4/80 positive cells in RPE-Choroid. Tissue extracts and flat-mounted retinas were prepared and evaluated by Western blot and by IHC. ( A ) Representative Western blot of RPE-Choroid homogenates prepared from WT mice without CNV, or 4 days after laser injury. Tubulin was used as loading control. Bands were quantified by densitometric analysis, and p75 NTR /tubulin ratio is represented in the bar graph expressed as units relative to control. Bars denote the mean ± SD from triplicate experiments, n = 5 mice/group. The asterisks show statistical differences respect to control. * p < 0.05. ( B ) Representative Western blot of RPE-Choroid homogenates prepared from WT mice without CNV, or 7 days after laser injury. Tubulin was used as loading control. Bands were quantified by densitometric analysis, and p75 NTR /tubulin ratio is represented in the bar graph expressed as units relative to control. ns: non-significant. Bars denote the mean ± SD from triplicate experiments, n = at least 4 mice/group. ( C – E ) Representative confocal images of RPE-Choroid flat-mounts of WT mice 4 days after laser injury, showing immunofluorescence staining with: ( C ) F4/80 (green), p75 NTR (red) and cell nuclei (blue). Scale bar upper panel: 50 µm. Scale bar lower panel: 20 µm. ( D ) IBA-1 (green), p75 NTR (red) and cell nuclei (blue). Scale bar upper panel: 100 µm. Scale bar lower panel: 50 µm. ( E ) CX3CR1 (green), p75 NTR (red) and cell nuclei (blue). Scale bar upper panel: 100 µm. Scale bar lower panel: 25 µm. ( F , G ) Representative confocal images of RPE-Choroid flat-mounts of WT mice 7 days after laser injury, showing immunofluorescence staining with ( F ) Isolectin IB-4 (green), p75 NTR (red) and cell nuclei (blue). Scale bar: 50 µm. ( G ) NG-2 (green), p75 NTR (red) and cell nuclei (blue). Scale bar: 50 µm.

Article Snippet: Afterwards, RPE-Choroids were incubated overnight with 0.01 μg/μL of Isolectin IB4 Alexa fluor-488 conjugate (GSA-IB4) from Molecular Probes, Inc. (Eugene, OR, USA), phalloidin-Alexa Fluor 647 (Thermo Fisher Scientific, A22287, Waltham, MA, USA), goat polyclonal anti-p75 NTR (1/500; R&D system), mouse anti f4/80 (1/50; Invitrogen, Waltham, MA, USA), rabbit anti CX3CR1 (1/100; Abcam, ab8021, Cambridge, UK), rabbit polyclonal anti-iba1 (1/200; Wako 019-19741), or rabbit polyclonal anti-NG-2 (1/100; AB5320, Millipore, Burlington, MA, USA).

Techniques: Expressing, Western Blot, Control, Immunofluorescence, Staining

Journal: Cells

Article Title: Etiological Roles of p75 NTR in a Mouse Model of Wet Age-Related Macular Degeneration

doi: 10.3390/cells12020297

Figure Lengend Snippet: Activated macroglial cells express p75 NTR after the laser around the injured area in the retina. p75 NTR protein co-localizes with GFAP-positive activated macroglia in CNV mice retinas, 7 days after laser. Tissue extracts and retina sections were prepared and evaluated by Western blot and by IHC. ( A ) Representative Western blot of total retinas homogenates prepared from WT mice without CNV, or 7 days after laser injury. (+) Correspond to a positive control (hippocampus E19 M2). Actin was used as loading control. Bands were quantified by densitometric analysis, and p75 NTR /actin ratio is represented in the bar graph expressed as units relative to control. Bars denote the mean ± SD from triplicate experiments, n = 6 mice/group. The asterisks show statistical differences respect to control. * p < 0.05. ( B ) Representative confocal images of WT CNV mice retinal cryosections, 7 days after laser. Upper panel: immunofluorescence staining of GFAP (green) and p75 NTR (red). Scale bar: 50 µm. * Indicates the injured area. Zoom scale bar: 25 µm. Cell nuclei counterstained with Hoechst are also shown (blue). Lower panel: immunofluorescence staining of β3 tubulin (green) and p75 NTR (red). Scale bar: 25 µm. Zoom scale bar: 10 µm. Abbreviations: GCL (ganglion cells layer), IPL (inner plexiform layer), INL (inner nuclear layer), OPL (outer plexiform layer), ONL (outer nuclear layer). ( C ) Representative confocal images of retinal flat-mount of WT CNV mice, 7 days after laser, showing immunofluorescence staining with NG-2 (green) and p75 NTR (red). Scale bar: 50 µm.

Article Snippet: Afterwards, RPE-Choroids were incubated overnight with 0.01 μg/μL of Isolectin IB4 Alexa fluor-488 conjugate (GSA-IB4) from Molecular Probes, Inc. (Eugene, OR, USA), phalloidin-Alexa Fluor 647 (Thermo Fisher Scientific, A22287, Waltham, MA, USA), goat polyclonal anti-p75 NTR (1/500; R&D system), mouse anti f4/80 (1/50; Invitrogen, Waltham, MA, USA), rabbit anti CX3CR1 (1/100; Abcam, ab8021, Cambridge, UK), rabbit polyclonal anti-iba1 (1/200; Wako 019-19741), or rabbit polyclonal anti-NG-2 (1/100; AB5320, Millipore, Burlington, MA, USA).

Techniques: Western Blot, Positive Control, Control, Immunofluorescence, Staining

Journal: Cells

Article Title: Etiological Roles of p75 NTR in a Mouse Model of Wet Age-Related Macular Degeneration

doi: 10.3390/cells12020297

Figure Lengend Snippet: Reduced inflammatory phenotype in the RPE-Choroids and retinas of p75 NTR knockout mice, after laser injury. Mononuclear phagocytic cell recruitment is significantly reduced in retinas and RPE-Choroids of p75 NTR KO mice after CNV. ( A ) Representative flow cytometry pseudocolor plots from WT and p75 NTR KO mice without CNV, or 4 days after laser injury. Cells in the gate were quantified and the number of cells in the gate/total cells ratio is represented in the bar graph expressed as units relative to WT no laser control. Graphs denote the mean ± SD from triplicate experiments, n = 6 mice/group. The asterisks show statistical differences respect to control. * p < 0.05, ** p < 0.01, *** p < 0.001. ( B ) Representative confocal images of RPE-Choroid flat-mounts of WT and p75 NTR KO mice 4 days after laser, showing immunofluorescence staining with Isolectin IB-4 (grey) and F4/80 (green). Scale bar: 200 µm. F4/80 fluorescence intensity and area were quantified with ImageJ FIJI software Version 1.53t, and represented in the bar graph expressed as units relative to CNV WT control. Bars denote the mean ± SD from triplicate experiments, n = at least 5 mice/group. ns: non-significant.

Article Snippet: Afterwards, RPE-Choroids were incubated overnight with 0.01 μg/μL of Isolectin IB4 Alexa fluor-488 conjugate (GSA-IB4) from Molecular Probes, Inc. (Eugene, OR, USA), phalloidin-Alexa Fluor 647 (Thermo Fisher Scientific, A22287, Waltham, MA, USA), goat polyclonal anti-p75 NTR (1/500; R&D system), mouse anti f4/80 (1/50; Invitrogen, Waltham, MA, USA), rabbit anti CX3CR1 (1/100; Abcam, ab8021, Cambridge, UK), rabbit polyclonal anti-iba1 (1/200; Wako 019-19741), or rabbit polyclonal anti-NG-2 (1/100; AB5320, Millipore, Burlington, MA, USA).

Techniques: Knock-Out, Flow Cytometry, Control, Immunofluorescence, Staining, Fluorescence, Software

Journal: Cells

Article Title: Etiological Roles of p75 NTR in a Mouse Model of Wet Age-Related Macular Degeneration

doi: 10.3390/cells12020297

Figure Lengend Snippet: Reduced neovascular phenotype in the choroid of p75 NTR knockout mice, after laser injury. The area and the perimeter of choroidal neovessels are significantly reduced in RPE-Choroid flat-mounts of p75 NTR KO mice. ( A ) Representative confocal images of RPE-Choroid flat-mounts of WT and p75 NTR KO mice 1 day after laser injury, showing immunofluorescence staining with Falloidin (green). The yellow outline represents the lesioned area estimated by F-actin negative staining. Scale bar: 15 µm. Cell nuclei counterstained with Hoechst are also shown (blue). The area and perimeter of the laser lesion were quantified with ImageJ FIJI software, and represented in the bar graph expressed as units relative to WT CNV control. Bars denote the mean ± SD from triplicate experiments, n = 4 mice/group. ns: non-significant. ( B ) Representative confocal images of RPE-Choroid flat-mounts of WT mice 7 days after laser injury, showing immunofluorescence staining with Isolectin IB-4 (green). The yellow outline represents the neovessels covered area. Scale bar: 200 µm. The area and the perimeter of the neovessels were quantified with ImageJ FIJI software, and represented in the bar graph expressed as units relative to CNV WT control. Bars denote the mean ± SD from triplicate experiments, n = at least 6 mice/group. The asterisks show statistical differences respect to control. * p < 0.05, ** p < 0.01, *** p < 0.001. ( C ) Representative Western blot of total retinal homogenates prepared from WT and p75 NTR KO mice without CNV, or 7 days after laser injury. Tubulin was used as loading control. Bands were quantified by densitometric analysis, and GFAP/tubulin ratio is represented in the bar graph expressed as units relative to control. Bars denote the mean ± SD from triplicate experiments, n = 3 mice/group. ( D ) Representative immunofluorescence analysis of GFAP (green) in retinal cryosections from WT and p75 NTR KO mice without injury or 7 days after laser injury. Scale bar: 50 µm. Cell nuclei counterstained with Hoechst are also shown (blue). Abbreviations: GCL (ganglion cells layer), IPL (inner plexiform layer), INL (inner nuclear layer), OPL (outer plexiform layer), ONL (outer nuclear layer). ( E ) Amplitudes and implicit times of a- and b-waves from scotopic electroretinograms recorded in WT and p75 NTR KO mice without injury or 7 days after laser injury. The data show averages of responses of both eyes. Graphs denote the mean ± SD from triplicate experiments, n = at least 6 mice/group. The symbol correspond: WT no laser (filled circle), WT CNV (filled square), p75 NTR KO no laser (filled triangle up), p75 NTR KO CNV (filled triangle down). The asterisks show statistical differences respect to control. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Afterwards, RPE-Choroids were incubated overnight with 0.01 μg/μL of Isolectin IB4 Alexa fluor-488 conjugate (GSA-IB4) from Molecular Probes, Inc. (Eugene, OR, USA), phalloidin-Alexa Fluor 647 (Thermo Fisher Scientific, A22287, Waltham, MA, USA), goat polyclonal anti-p75 NTR (1/500; R&D system), mouse anti f4/80 (1/50; Invitrogen, Waltham, MA, USA), rabbit anti CX3CR1 (1/100; Abcam, ab8021, Cambridge, UK), rabbit polyclonal anti-iba1 (1/200; Wako 019-19741), or rabbit polyclonal anti-NG-2 (1/100; AB5320, Millipore, Burlington, MA, USA).

Techniques: Knock-Out, Immunofluorescence, Staining, Negative Staining, Software, Control, Western Blot

Journal: Cells

Article Title: Etiological Roles of p75 NTR in a Mouse Model of Wet Age-Related Macular Degeneration

doi: 10.3390/cells12020297

Figure Lengend Snippet: Reduced neovascular phenotype and improved retinal function in p75 NTR antagonist-treated wild type mice after laser injury. p75 NTR receptor antagonist reduced the area and perimeter of choroidal neovessels and improved retinal functionality in WT CNV mice. ( A ) Representative confocal images of RPE-Choroid flat-mounts of WT mice 7 days after laser, showing immunofluorescence staining with Isolectin IB-4 (grey). Scale bar: 100 µm. ( B ) The area and perimeter of neovessels were quantified with ImageJ FIJI software and represented in the bar graph expressed as units relative to WT CNV vehicle control. Bars denote the mean ± SD from triplicate experiments, n = 5 mice/group. ( C ) Amplitudes and implicit times of a- and b-waves from scotopic electroretinograms recorded 7 days after the laser in WT and WT CNV mice injected with THX-B or vehicle. The data show averages of responses of both. Graphs denote the mean ± SD from triplicate experiments, n = at least 6 mice/group. ( D ) Representative flow cytometry pseudocolor plots from WT CNV and no laser mice injected with THX-B or vehicle, 4 days after laser. Cells in the gate were quantified and the number of cells in the gate/total cells ratio is represented in the bar graph expressed as units relative to WT no laser vehicle control. Graphs denote the mean ± SD from triplicate experiments, n = 6 mice/group. The symbol correspond: No laser (white circle), CNV Vehicle (filled square), No laser THX B (filled triangle up), CNV THX B (filled triangle down). The asterisks show statistical differences respect to control. * p < 0.05, ** p < 0.01, *** p < 0.001. ( E ) Representative confocal images of RPE-Choroid flat-mounts of WT mice 4 days after laser, showing immunofluorescence staining with F4/80 (green). Scale bar: 200 µm. The area and the F4/80 fluorescence intensity were quantified with ImageJ FIJI software and represented in the bar graph expressed as units relative to WT CNV+ vehicle control. Bars denote the mean ± SD from triplicate experiments, n = 4 mice/group. ns: non-significant.

Article Snippet: Afterwards, RPE-Choroids were incubated overnight with 0.01 μg/μL of Isolectin IB4 Alexa fluor-488 conjugate (GSA-IB4) from Molecular Probes, Inc. (Eugene, OR, USA), phalloidin-Alexa Fluor 647 (Thermo Fisher Scientific, A22287, Waltham, MA, USA), goat polyclonal anti-p75 NTR (1/500; R&D system), mouse anti f4/80 (1/50; Invitrogen, Waltham, MA, USA), rabbit anti CX3CR1 (1/100; Abcam, ab8021, Cambridge, UK), rabbit polyclonal anti-iba1 (1/200; Wako 019-19741), or rabbit polyclonal anti-NG-2 (1/100; AB5320, Millipore, Burlington, MA, USA).

Techniques: Immunofluorescence, Staining, Software, Control, Injection, Flow Cytometry, Fluorescence

Journal: iScience

Article Title: Cholinotrophic basal forebrain connectome dysfunction in Down syndrome with and without dementia

doi: 10.1016/j.isci.2025.113041

Figure Lengend Snippet: Images and quantitation of cholinotrophic basal forebrain neuron pathology within the nucleus basalis of Meynert in DSD+, DSD- and AMC cases (A–N) Photomicrographs show p75 NTR (brown, A-C), ChAT (purple, D-F) positive cells in AMC, DSD-, and DSD+ cases. Note the decrease in the number and intensity of both p75 NTR and ChAT labeled neurons in DSD- cases compared to an even greater reduction in DSD+ and globose shaped cells (black arrows) in DSD+ cases (C, F). Arrows in (A–F) mark cells shown at a higher magnification in the boxed areas located at the lower right corner of each panel. Dark brown AT8 (G, I) and TauC3 (H, J) bearing neurofibrillary tangles (NFTs) were observed only in the DSD- and DSD+ cases. Black arrows indicate globose shaped NFTs (G, H, and I) that are shown at a higher magnification in boxed areas adjacent to the lower magnification images. Note that not all neurons within the nbM (thin black arrows) contained tau pathology (G, H) in DSD- cases. Moreover, TauC3 staining revealed two NFT phenotypes that displayed either peripherally located or intense labeling that filled the entire structure (see boxed images adjacent to (H) and (J)). Scale bar in F = 50 μm and inset = 20 μm applies to panels A-E. Scale bar in J = 20 μm and inset = 20 μm applies to (G–J). Histograms show a significant reduction in both p75 NTR (K) and ChAT (L) positive cells in DSD+ compared to AMC. Although no significant were found in number of AT8 or TauC3 NFT positive cells (M), there was an increase in NTs in DSD+ compared to DSD- (N). ACM n = 5, DSD- n = 5, DSD+ n = 10. Data shown are presented as mean ± SEM. Statistical significance was determined using the Kruskal–Wallis’s test followed by Dunn’s test for comparisons across three clinical groups, and the Mann–Whitney test for comparisons between two groups (DSD- vs. DSD+). Significance levels (∗) were set at: ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Antibodies were added to blocking buffer and membranes incubated overnight (4°C) in mouse anti-p75 NTR ([1:150], Santa Cruz Biotechnology Cat# sc-271708, RRID: AB_10714958 ), rabbit anti-ChAT ([1:500] (Proteintech Cat# 20747-1-AP, RRID: AB_10898169 ), rabbit anti-TrkA ([1:150], Fitzgerald Industries International Cat# 20R-TR013, RRID: AB_1289098 ) or rabbit anti-proNGF ([1:250], Alomone Labs Cat# ANT-005, RRID: AB_2040021 ), washed, incubated with horseradish peroxidase-conjugated with either a goat anti-mouse IgG (1:200) or goat anti-rabbit IgG (1:200) secondary antibody at RT, visualized by chemiluminescence (Kodak Image Station 440CF; Perkin-Elmer, Wellesley, MA), and quantified with Kodak 1.

Techniques: Quantitation Assay, Labeling, Staining, MANN-WHITNEY

Journal: iScience

Article Title: Cholinotrophic basal forebrain connectome dysfunction in Down syndrome with and without dementia

doi: 10.1016/j.isci.2025.113041

Figure Lengend Snippet: Immunofluorescent images and quantification of p75 NTR neurons with and without AT8 and MAP2, as well as Thioflavin S single and AT8/Thioflavin S dual-labeled nbM cells in DSD+ and DSD-cases. (A–U) Immunofluorescent neurons labeled with p75 NTR (A, E, green), MAP2 (B, F, red), AT8 (C, G, Cyan), ThS (blue) (I, L) and merge images (D, H, K, N, R) in DSD- and DSD+ cases. In the DSD-cases there were greater p75 NTR cells AT8 negative neurons and p75 NTR MAP2 dual labeled neurons (A-D) compared to the DSD+ cases (E–H). Note that not all p75 NTR cells colocalize with AT8 or MAP2 in both DS groups (D, H). To determine the stage of a tangle tissue was stained for ThS, a marker of advanced pathology and the early stage AT8 phosphorylation antibody. Note that there are only a few ThS-labeled tangles that displayed AT8 in both DS groups (yellow arrows), compared to single ThS tangles, which were greater in DSD+ than in DSD- (white arrows) (I-L). Note that ThS positive NFTs that do not contain AT8 also do not colocalize with MAP2 (O-R). Scale bar in F = 25 μm applies to panels A-G, N = 25 μm applies to panels I-M and R = 10 μm and applies to O-Q. Graph showing a significant reduction in both p75 NTR AT8 immuno-negative, and p75 NTR MAP2 dual labeled neurons in DSD+ compared to DSD- (S). ThS-positive cells were greater (T), while the percentage of double-labeled cells with AT8 and ThS decreased (U) in DSD+ compared to individuals without dementia. DSD- n = 5, DSD+ n = 5. Data are presented as mean ± SEM. Statistical significance was determined using Mann–Whitney test for comparisons between DSD- and DSD+. Significance levels (∗) were set at: ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: Antibodies were added to blocking buffer and membranes incubated overnight (4°C) in mouse anti-p75 NTR ([1:150], Santa Cruz Biotechnology Cat# sc-271708, RRID: AB_10714958 ), rabbit anti-ChAT ([1:500] (Proteintech Cat# 20747-1-AP, RRID: AB_10898169 ), rabbit anti-TrkA ([1:150], Fitzgerald Industries International Cat# 20R-TR013, RRID: AB_1289098 ) or rabbit anti-proNGF ([1:250], Alomone Labs Cat# ANT-005, RRID: AB_2040021 ), washed, incubated with horseradish peroxidase-conjugated with either a goat anti-mouse IgG (1:200) or goat anti-rabbit IgG (1:200) secondary antibody at RT, visualized by chemiluminescence (Kodak Image Station 440CF; Perkin-Elmer, Wellesley, MA), and quantified with Kodak 1.

Techniques: Labeling, Staining, Marker, Phospho-proteomics, MANN-WHITNEY

Journal: iScience

Article Title: Cholinotrophic basal forebrain connectome dysfunction in Down syndrome with and without dementia

doi: 10.1016/j.isci.2025.113041

Figure Lengend Snippet: FC cholinotrophic protein levels in AMC, DSD- and DSD+ (A–D) Representative immunoblots, and bar graphs show a significant upregulation of (A) proNGF and (B) p75 NTR , while (C) ChAT protein was downregulated between AMC and DSD+. (D) TrkA protein levels were stable across the groups analyzed. ACM n = 5, DSD- n = 5, DSD+ n = 13. Data are presented as mean ± SEM. Statistical significance was determined using the Kruskal–Wallis’s test followed by Dunn’s test for comparisons across three clinical groups. Significance levels (∗) were set at: ∗ p < 0.05.

Article Snippet: Antibodies were added to blocking buffer and membranes incubated overnight (4°C) in mouse anti-p75 NTR ([1:150], Santa Cruz Biotechnology Cat# sc-271708, RRID: AB_10714958 ), rabbit anti-ChAT ([1:500] (Proteintech Cat# 20747-1-AP, RRID: AB_10898169 ), rabbit anti-TrkA ([1:150], Fitzgerald Industries International Cat# 20R-TR013, RRID: AB_1289098 ) or rabbit anti-proNGF ([1:250], Alomone Labs Cat# ANT-005, RRID: AB_2040021 ), washed, incubated with horseradish peroxidase-conjugated with either a goat anti-mouse IgG (1:200) or goat anti-rabbit IgG (1:200) secondary antibody at RT, visualized by chemiluminescence (Kodak Image Station 440CF; Perkin-Elmer, Wellesley, MA), and quantified with Kodak 1.

Techniques: Western Blot

Journal: iScience

Article Title: Cholinotrophic basal forebrain connectome dysfunction in Down syndrome with and without dementia

doi: 10.1016/j.isci.2025.113041

Figure Lengend Snippet: Summary of changes in the cholinotrophic basal forebrain connectome in DS Diagrammatic sagittal view of the human brain (A) and a modified stacked bar graph (B) illustrating differences in the pathobiology of the cholinotrophic projection system between non-trisomy age-matched control (AMC), DS without dementia (DSD-) and DS with dementia (DSD+) individuals with DS. Frontal cortex protein levels for ChAT (pink), proNGF (green), p75 NTR (purple) and TrkA (light blue). Nucleus basalis NFTs of ThS (blue), AT8 (orange) and TauC3 (black) and counts of p75 NTR and ChAT (red) neurons. Created with BioRender.com .

Article Snippet: Antibodies were added to blocking buffer and membranes incubated overnight (4°C) in mouse anti-p75 NTR ([1:150], Santa Cruz Biotechnology Cat# sc-271708, RRID: AB_10714958 ), rabbit anti-ChAT ([1:500] (Proteintech Cat# 20747-1-AP, RRID: AB_10898169 ), rabbit anti-TrkA ([1:150], Fitzgerald Industries International Cat# 20R-TR013, RRID: AB_1289098 ) or rabbit anti-proNGF ([1:250], Alomone Labs Cat# ANT-005, RRID: AB_2040021 ), washed, incubated with horseradish peroxidase-conjugated with either a goat anti-mouse IgG (1:200) or goat anti-rabbit IgG (1:200) secondary antibody at RT, visualized by chemiluminescence (Kodak Image Station 440CF; Perkin-Elmer, Wellesley, MA), and quantified with Kodak 1.

Techniques: Modification, Control

Journal: Materials Today Bio

Article Title: Fabrication of GDNF-Gel/HA-Mg nerve conduit and its role in repairing peripheral nerve defects

doi: 10.1016/j.mtbio.2025.101764

Figure Lengend Snippet: GDNF-Gel/HA-Mg extracts on NGF secretion, p75 NTR and Schwann cell proliferation, migration related genes expression. A: ELISA was used to detect the concentration of NGF in the supernatant. BC: The effect of different concentrations of extracts on the expression level of the p75NTR in Schwann cells detected by western blotting and statistical analysis; DE: The effect of different concentrations of extracts on the expression level of the p75NTR in Schwann cells detected by immunofluorescence and statistical analysis; F–J: the expression of Netrin-1, Rac1, Cdc42 and UNC5b in Schwann cells detected by western blotting and statistical analysis. NGF: nerve growth factor; p75 NTR : p75 neurotrophin receptor; IOD: integrated optical density; ns: no statistical significance; ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

Article Snippet: Blocked with 5 % defatted milk for 2 h, washed three times with TBST, and incubated with anti-p75 NTR (1:2000) (Proteintech, China), Netrin-1 (1:1000, Selleck, USA), Rac1 (1:1000, Selleck, USA), Cdc42 (1:1000, Selleck, USA), UNC5b (1:500, Selleck, USA) antibody overnight at 4 °C.

Techniques: Migration, Expressing, Enzyme-linked Immunosorbent Assay, Concentration Assay, Western Blot, Immunofluorescence

Journal: Materials Today Bio

Article Title: Fabrication of GDNF-Gel/HA-Mg nerve conduit and its role in repairing peripheral nerve defects

doi: 10.1016/j.mtbio.2025.101764

Figure Lengend Snippet: The effect of GDNF-Gel/HA-Mg conduit on myelination of regenerated sciatic nerves and prevention of gastrocnemius muscle atrophy. AB: Toluidine blue staining results of sciatic nerves in different groups and statistical analysis; CD: The expression of p75 NTR detected by immunofluorescence and statistical analysis; EF: The morphologies of gastrocnemius muscle in different groups and statistical analysis. Mg: magnesium; HA: hydroxylapatite; GDNF: glial cell line-derived neurotrophic factor; GelMA: Gelatin methacryloyl; p75 NTR : p75 neurotrophin receptor. ns: no statistical significance; ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Blocked with 5 % defatted milk for 2 h, washed three times with TBST, and incubated with anti-p75 NTR (1:2000) (Proteintech, China), Netrin-1 (1:1000, Selleck, USA), Rac1 (1:1000, Selleck, USA), Cdc42 (1:1000, Selleck, USA), UNC5b (1:500, Selleck, USA) antibody overnight at 4 °C.

Techniques: Staining, Expressing, Immunofluorescence, Derivative Assay

Journal: Cell Death & Disease

Article Title: AraC interacts with p75 NTR transmembrane domain to induce cell death of mature neurons

doi: 10.1038/s41419-023-05979-7

Figure Lengend Snippet: A Representative micrographs of wild-type P7 CGNs cultured for 4 DIV and double stained with anti-p75 NTR together with anti- β III tubulin and counterstained with DAPI. Scale bars, 50 μm. B Representative and quantification of immunoblots showing the expression of p75 NTR in 4DIV CGNs treated with PBS (0 μM AraC), 50 μM or 500 μM AraC for 24 h. C Image of representative P7 p75 NTR+/+ and p75 NTR−/− CGNs cultured for 4 DIV, treated with either PBS (control) or 500 μM AraC for 24 h and stained for anti-cleaved caspase 3 (magenta), TUNEL (green) and counterstained with DAPI (blue). Scale bars, 50 μm. D , E Quantification of percentage cleaved caspase 3 positive ( D ) and TUNEL positive ( E ) in p75 NTR+/+ and p75 NTR−/− neurons treated with PBS or AraC (500 μM or 1000 μM for 24 h (total of 80 images per condition were counted)). Mean ± s.e.m. of data from four separate cultures, * P < 0.05 and ** P < 0.01 compared to control, one-way ANOVA followed by Bonferroni post hoc test) is shown.

Article Snippet: The primary antibodies used in this study were: polyclonal anti-cleaved caspase 3 (Cell Signalling Technology; 9761; 1:400), monoclonal anti–β-III tubulin (R&D systems, Cat: MAB1195, 1:10000), polyclonal anti-p75 NTR (Neuromics; GT15057; 1:500), polyclonal anti-TrkB (R&D systems; AF1494; 1:250), polyclonal anti-MAP2 (Abcam; ab5392; 1:2000) and polyclonal anti-P65NFkB (Santa Cruz; sc-372; 1:250).

Techniques: Cell Culture, Staining, Western Blot, Expressing, TUNEL Assay

Journal: Cell Death & Disease

Article Title: AraC interacts with p75 NTR transmembrane domain to induce cell death of mature neurons

doi: 10.1038/s41419-023-05979-7

Figure Lengend Snippet: A Ranked list of potential AraC binding sites and their distance from the highest scoring model. B Three principal AraC binding sites were identified and are shown in blue, green and orange stick representation docked onto p75 NTR TMD. p75 NTR TMD is shown in tan with a transparent surface, and residues potentially interacting with docked AraC are shown in stick representation. C 2D interaction diagrams of the three respective modes.

Article Snippet: The primary antibodies used in this study were: polyclonal anti-cleaved caspase 3 (Cell Signalling Technology; 9761; 1:400), monoclonal anti–β-III tubulin (R&D systems, Cat: MAB1195, 1:10000), polyclonal anti-p75 NTR (Neuromics; GT15057; 1:500), polyclonal anti-TrkB (R&D systems; AF1494; 1:250), polyclonal anti-MAP2 (Abcam; ab5392; 1:2000) and polyclonal anti-P65NFkB (Santa Cruz; sc-372; 1:250).

Techniques: Binding Assay

Journal: Cell Death & Disease

Article Title: AraC interacts with p75 NTR transmembrane domain to induce cell death of mature neurons

doi: 10.1038/s41419-023-05979-7

Figure Lengend Snippet: A Dose response of AraC in the AraTM assay of p75 NTR . Results are plotted as means ± SD ( N = 3). B – D Comparison of human wild-type p75 NTR TMD with p75 NTR TMD with I252A plus V254A ( B ) or P253G ( C ) or C256A ( D ) mutants in the AraTM assay in response to increasing doses of AraC. The GFP over OD630 signal without any drug added was set at 100%. Results are plotted as means ± SD ( N = 3). Significance was calculated using the two-way ANOVA followed by Bonferroni post hoc test where ** p < 0.01 and *** p < 0.001.

Article Snippet: The primary antibodies used in this study were: polyclonal anti-cleaved caspase 3 (Cell Signalling Technology; 9761; 1:400), monoclonal anti–β-III tubulin (R&D systems, Cat: MAB1195, 1:10000), polyclonal anti-p75 NTR (Neuromics; GT15057; 1:500), polyclonal anti-TrkB (R&D systems; AF1494; 1:250), polyclonal anti-MAP2 (Abcam; ab5392; 1:2000) and polyclonal anti-P65NFkB (Santa Cruz; sc-372; 1:250).

Techniques: AraTM Assay

Journal: Cell Death & Disease

Article Title: AraC interacts with p75 NTR transmembrane domain to induce cell death of mature neurons

doi: 10.1038/s41419-023-05979-7

Figure Lengend Snippet: A Representative immunoblot probed for p75 NTR and GAPDH and quantification of CETSA for p75 NTR protein response to different temperatures. B Representative immunoblot probed for p75 NTR and GAPDH and quantification of p75 NTR CETSA for AraC dose–response on protein lysate extracted from HEK293 cells constitutively overexpressing p75 NTR . The lysate was heated to 37 °C and 53 °C. C Representative immunoblot probed for p75 NTR and GAPDH and quantification of p75 NTR CETSA for p75 NTR response to temperature after treatment with either PBS (vehicle) or 500 μM AraC. Mean ± s.e.m. of data from 3 to 6 separate cultures is shown for p75 NTR . D Live cell homo-FRET anisotropy of p75 NTR in COS-7 cells in response to AraC. Shown are representative time-lapse images before (−3 min) and after (0.5, 4 and 9 min) addition of 500 µM AraC or DMSO (control). Scale bars, 5 μm. E Live cell homo-FRET anisotropy of p75 NTR in COS-7 cells in response to AraC. Shown are representative traces of average anisotropy change after the addition of AraC (500 µM at 0 min) or vehicle in cells expressing wild-type rat p75 NTR . F Integrated peak area of live cell homo-FRET anisotropy of p75 NTR in COS-7 cells in response to AraC (considering the area under and above y = 1). Results are plotted as means ± SD ( N = 7). *** P < 0.001; Student t -test. G Oscillations/min of live cell homo-FRET anisotropy of p75 NTR in COS7 cells in response to AraC (considering the vehicle oscillations as threshold). Results are plotted as means ± SD ( N = 7).

Article Snippet: The primary antibodies used in this study were: polyclonal anti-cleaved caspase 3 (Cell Signalling Technology; 9761; 1:400), monoclonal anti–β-III tubulin (R&D systems, Cat: MAB1195, 1:10000), polyclonal anti-p75 NTR (Neuromics; GT15057; 1:500), polyclonal anti-TrkB (R&D systems; AF1494; 1:250), polyclonal anti-MAP2 (Abcam; ab5392; 1:2000) and polyclonal anti-P65NFkB (Santa Cruz; sc-372; 1:250).

Techniques: Western Blot, Expressing

Journal: Cell Death & Disease

Article Title: AraC interacts with p75 NTR transmembrane domain to induce cell death of mature neurons

doi: 10.1038/s41419-023-05979-7

Figure Lengend Snippet: A Representative micrographs of wild-type P7 CGNs cultured for 4DIV and double stained with anti-TRAF6 together with anti- β III tubulin and counterstained with DAPI. Scale bars, 50 μm. B Micrographs of p75 NTR :TRAF6 PLA (red) in CGNs treated with either PBS (control) or 500 μM AraC for 10 min. Images were selected from 25 images per condition from 3 to 4 separate experiments. Scale bar, 20 μm. C Quantification of p75 NTR :TRAF6 PLA puncta in CGNs treated with either PBS (control) or 500 μM AraC for 10 min. Mean ± sem of data from 3 experiments (* P < 0.05; unpaired Student t -test) is shown. D Representative western blots probed with phospho-c-Jun (Thr91), total c-Jun and GAPDH of lysates of wild type P7 CGNs grown for 4 days prior to 15-, 30- or 60-min treatment with 500 μM AraC. D Quantification of c-Jun (Thr91) phosphorylation of total lysate of untreated wild-type P7 CGNs or neurons treated with 500 μM for 15, 30 and 60 min. Mean ± sem of densitometry from 4 experiments (* P < 0.05; one-way ANOVA followed by Bonferroni test) is shown.

Article Snippet: The primary antibodies used in this study were: polyclonal anti-cleaved caspase 3 (Cell Signalling Technology; 9761; 1:400), monoclonal anti–β-III tubulin (R&D systems, Cat: MAB1195, 1:10000), polyclonal anti-p75 NTR (Neuromics; GT15057; 1:500), polyclonal anti-TrkB (R&D systems; AF1494; 1:250), polyclonal anti-MAP2 (Abcam; ab5392; 1:2000) and polyclonal anti-P65NFkB (Santa Cruz; sc-372; 1:250).

Techniques: Cell Culture, Staining, Western Blot

Journal: Cell Death & Disease

Article Title: AraC interacts with p75 NTR transmembrane domain to induce cell death of mature neurons

doi: 10.1038/s41419-023-05979-7

Figure Lengend Snippet: This schematic diagram depicts p75 NTR signalling in healthy mature neurons and when AraC interacts with P75 NTR . Upon AraC binding to P75 NTR , RIPK2 is dislodged from the death domain (DD), allowing the exposure of the juxta membrane (JX), where TRAF6 binds. The binding of TRAF6 to p75 NTR activates the JNK pathways that lead to the translocation of c-Jun to the nucleus and transcription of pro-apoptotic genes. Figure produced in BioRender.

Article Snippet: The primary antibodies used in this study were: polyclonal anti-cleaved caspase 3 (Cell Signalling Technology; 9761; 1:400), monoclonal anti–β-III tubulin (R&D systems, Cat: MAB1195, 1:10000), polyclonal anti-p75 NTR (Neuromics; GT15057; 1:500), polyclonal anti-TrkB (R&D systems; AF1494; 1:250), polyclonal anti-MAP2 (Abcam; ab5392; 1:2000) and polyclonal anti-P65NFkB (Santa Cruz; sc-372; 1:250).

Techniques: Binding Assay, Translocation Assay, Produced

Journal: Cells

Article Title: Etiological Roles of p75 NTR in a Mouse Model of Wet Age-Related Macular Degeneration

doi: 10.3390/cells12020297

Figure Lengend Snippet: MPCs expressing p75 NTR migrate into the laser-injured area of RPE-Choroid. p75 NTR co-localizes in F4/80 positive cells in RPE-Choroid. Tissue extracts and flat-mounted retinas were prepared and evaluated by Western blot and by IHC. ( A ) Representative Western blot of RPE-Choroid homogenates prepared from WT mice without CNV, or 4 days after laser injury. Tubulin was used as loading control. Bands were quantified by densitometric analysis, and p75 NTR /tubulin ratio is represented in the bar graph expressed as units relative to control. Bars denote the mean ± SD from triplicate experiments, n = 5 mice/group. The asterisks show statistical differences respect to control. * p < 0.05. ( B ) Representative Western blot of RPE-Choroid homogenates prepared from WT mice without CNV, or 7 days after laser injury. Tubulin was used as loading control. Bands were quantified by densitometric analysis, and p75 NTR /tubulin ratio is represented in the bar graph expressed as units relative to control. ns: non-significant. Bars denote the mean ± SD from triplicate experiments, n = at least 4 mice/group. ( C – E ) Representative confocal images of RPE-Choroid flat-mounts of WT mice 4 days after laser injury, showing immunofluorescence staining with: ( C ) F4/80 (green), p75 NTR (red) and cell nuclei (blue). Scale bar upper panel: 50 µm. Scale bar lower panel: 20 µm. ( D ) IBA-1 (green), p75 NTR (red) and cell nuclei (blue). Scale bar upper panel: 100 µm. Scale bar lower panel: 50 µm. ( E ) CX3CR1 (green), p75 NTR (red) and cell nuclei (blue). Scale bar upper panel: 100 µm. Scale bar lower panel: 25 µm. ( F , G ) Representative confocal images of RPE-Choroid flat-mounts of WT mice 7 days after laser injury, showing immunofluorescence staining with ( F ) Isolectin IB-4 (green), p75 NTR (red) and cell nuclei (blue). Scale bar: 50 µm. ( G ) NG-2 (green), p75 NTR (red) and cell nuclei (blue). Scale bar: 50 µm.

Article Snippet: The following antibodies were used: rabbit polyclonal anti-p75 NTR (PRB-602C Covance, now Biolegend), rabbit polyclonal anti-GFAP (1/1000; Dako, Carpinteria, CA, USA), mouse monoclonal anti-β tubulin (1/5000; Sigma), mouse monoclonal anti-β-actin (1/2000; ab8226, Abcam).

Techniques: Expressing, Western Blot, Control, Immunofluorescence, Staining

Journal: Cells

Article Title: Etiological Roles of p75 NTR in a Mouse Model of Wet Age-Related Macular Degeneration

doi: 10.3390/cells12020297

Figure Lengend Snippet: Activated macroglial cells express p75 NTR after the laser around the injured area in the retina. p75 NTR protein co-localizes with GFAP-positive activated macroglia in CNV mice retinas, 7 days after laser. Tissue extracts and retina sections were prepared and evaluated by Western blot and by IHC. ( A ) Representative Western blot of total retinas homogenates prepared from WT mice without CNV, or 7 days after laser injury. (+) Correspond to a positive control (hippocampus E19 M2). Actin was used as loading control. Bands were quantified by densitometric analysis, and p75 NTR /actin ratio is represented in the bar graph expressed as units relative to control. Bars denote the mean ± SD from triplicate experiments, n = 6 mice/group. The asterisks show statistical differences respect to control. * p < 0.05. ( B ) Representative confocal images of WT CNV mice retinal cryosections, 7 days after laser. Upper panel: immunofluorescence staining of GFAP (green) and p75 NTR (red). Scale bar: 50 µm. * Indicates the injured area. Zoom scale bar: 25 µm. Cell nuclei counterstained with Hoechst are also shown (blue). Lower panel: immunofluorescence staining of β3 tubulin (green) and p75 NTR (red). Scale bar: 25 µm. Zoom scale bar: 10 µm. Abbreviations: GCL (ganglion cells layer), IPL (inner plexiform layer), INL (inner nuclear layer), OPL (outer plexiform layer), ONL (outer nuclear layer). ( C ) Representative confocal images of retinal flat-mount of WT CNV mice, 7 days after laser, showing immunofluorescence staining with NG-2 (green) and p75 NTR (red). Scale bar: 50 µm.

Article Snippet: The following antibodies were used: rabbit polyclonal anti-p75 NTR (PRB-602C Covance, now Biolegend), rabbit polyclonal anti-GFAP (1/1000; Dako, Carpinteria, CA, USA), mouse monoclonal anti-β tubulin (1/5000; Sigma), mouse monoclonal anti-β-actin (1/2000; ab8226, Abcam).

Techniques: Western Blot, Positive Control, Control, Immunofluorescence, Staining

Journal: Cells

Article Title: Etiological Roles of p75 NTR in a Mouse Model of Wet Age-Related Macular Degeneration

doi: 10.3390/cells12020297

Figure Lengend Snippet: Reduced inflammatory phenotype in the RPE-Choroids and retinas of p75 NTR knockout mice, after laser injury. Mononuclear phagocytic cell recruitment is significantly reduced in retinas and RPE-Choroids of p75 NTR KO mice after CNV. ( A ) Representative flow cytometry pseudocolor plots from WT and p75 NTR KO mice without CNV, or 4 days after laser injury. Cells in the gate were quantified and the number of cells in the gate/total cells ratio is represented in the bar graph expressed as units relative to WT no laser control. Graphs denote the mean ± SD from triplicate experiments, n = 6 mice/group. The asterisks show statistical differences respect to control. * p < 0.05, ** p < 0.01, *** p < 0.001. ( B ) Representative confocal images of RPE-Choroid flat-mounts of WT and p75 NTR KO mice 4 days after laser, showing immunofluorescence staining with Isolectin IB-4 (grey) and F4/80 (green). Scale bar: 200 µm. F4/80 fluorescence intensity and area were quantified with ImageJ FIJI software Version 1.53t, and represented in the bar graph expressed as units relative to CNV WT control. Bars denote the mean ± SD from triplicate experiments, n = at least 5 mice/group. ns: non-significant.

Article Snippet: The following antibodies were used: rabbit polyclonal anti-p75 NTR (PRB-602C Covance, now Biolegend), rabbit polyclonal anti-GFAP (1/1000; Dako, Carpinteria, CA, USA), mouse monoclonal anti-β tubulin (1/5000; Sigma), mouse monoclonal anti-β-actin (1/2000; ab8226, Abcam).

Techniques: Knock-Out, Flow Cytometry, Control, Immunofluorescence, Staining, Fluorescence, Software

Journal: Cells

Article Title: Etiological Roles of p75 NTR in a Mouse Model of Wet Age-Related Macular Degeneration

doi: 10.3390/cells12020297

Figure Lengend Snippet: Reduced neovascular phenotype in the choroid of p75 NTR knockout mice, after laser injury. The area and the perimeter of choroidal neovessels are significantly reduced in RPE-Choroid flat-mounts of p75 NTR KO mice. ( A ) Representative confocal images of RPE-Choroid flat-mounts of WT and p75 NTR KO mice 1 day after laser injury, showing immunofluorescence staining with Falloidin (green). The yellow outline represents the lesioned area estimated by F-actin negative staining. Scale bar: 15 µm. Cell nuclei counterstained with Hoechst are also shown (blue). The area and perimeter of the laser lesion were quantified with ImageJ FIJI software, and represented in the bar graph expressed as units relative to WT CNV control. Bars denote the mean ± SD from triplicate experiments, n = 4 mice/group. ns: non-significant. ( B ) Representative confocal images of RPE-Choroid flat-mounts of WT mice 7 days after laser injury, showing immunofluorescence staining with Isolectin IB-4 (green). The yellow outline represents the neovessels covered area. Scale bar: 200 µm. The area and the perimeter of the neovessels were quantified with ImageJ FIJI software, and represented in the bar graph expressed as units relative to CNV WT control. Bars denote the mean ± SD from triplicate experiments, n = at least 6 mice/group. The asterisks show statistical differences respect to control. * p < 0.05, ** p < 0.01, *** p < 0.001. ( C ) Representative Western blot of total retinal homogenates prepared from WT and p75 NTR KO mice without CNV, or 7 days after laser injury. Tubulin was used as loading control. Bands were quantified by densitometric analysis, and GFAP/tubulin ratio is represented in the bar graph expressed as units relative to control. Bars denote the mean ± SD from triplicate experiments, n = 3 mice/group. ( D ) Representative immunofluorescence analysis of GFAP (green) in retinal cryosections from WT and p75 NTR KO mice without injury or 7 days after laser injury. Scale bar: 50 µm. Cell nuclei counterstained with Hoechst are also shown (blue). Abbreviations: GCL (ganglion cells layer), IPL (inner plexiform layer), INL (inner nuclear layer), OPL (outer plexiform layer), ONL (outer nuclear layer). ( E ) Amplitudes and implicit times of a- and b-waves from scotopic electroretinograms recorded in WT and p75 NTR KO mice without injury or 7 days after laser injury. The data show averages of responses of both eyes. Graphs denote the mean ± SD from triplicate experiments, n = at least 6 mice/group. The symbol correspond: WT no laser (filled circle), WT CNV (filled square), p75 NTR KO no laser (filled triangle up), p75 NTR KO CNV (filled triangle down). The asterisks show statistical differences respect to control. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The following antibodies were used: rabbit polyclonal anti-p75 NTR (PRB-602C Covance, now Biolegend), rabbit polyclonal anti-GFAP (1/1000; Dako, Carpinteria, CA, USA), mouse monoclonal anti-β tubulin (1/5000; Sigma), mouse monoclonal anti-β-actin (1/2000; ab8226, Abcam).

Techniques: Knock-Out, Immunofluorescence, Staining, Negative Staining, Software, Control, Western Blot

Journal: Cells

Article Title: Etiological Roles of p75 NTR in a Mouse Model of Wet Age-Related Macular Degeneration

doi: 10.3390/cells12020297

Figure Lengend Snippet: Reduced neovascular phenotype and improved retinal function in p75 NTR antagonist-treated wild type mice after laser injury. p75 NTR receptor antagonist reduced the area and perimeter of choroidal neovessels and improved retinal functionality in WT CNV mice. ( A ) Representative confocal images of RPE-Choroid flat-mounts of WT mice 7 days after laser, showing immunofluorescence staining with Isolectin IB-4 (grey). Scale bar: 100 µm. ( B ) The area and perimeter of neovessels were quantified with ImageJ FIJI software and represented in the bar graph expressed as units relative to WT CNV vehicle control. Bars denote the mean ± SD from triplicate experiments, n = 5 mice/group. ( C ) Amplitudes and implicit times of a- and b-waves from scotopic electroretinograms recorded 7 days after the laser in WT and WT CNV mice injected with THX-B or vehicle. The data show averages of responses of both. Graphs denote the mean ± SD from triplicate experiments, n = at least 6 mice/group. ( D ) Representative flow cytometry pseudocolor plots from WT CNV and no laser mice injected with THX-B or vehicle, 4 days after laser. Cells in the gate were quantified and the number of cells in the gate/total cells ratio is represented in the bar graph expressed as units relative to WT no laser vehicle control. Graphs denote the mean ± SD from triplicate experiments, n = 6 mice/group. The symbol correspond: No laser (white circle), CNV Vehicle (filled square), No laser THX B (filled triangle up), CNV THX B (filled triangle down). The asterisks show statistical differences respect to control. * p < 0.05, ** p < 0.01, *** p < 0.001. ( E ) Representative confocal images of RPE-Choroid flat-mounts of WT mice 4 days after laser, showing immunofluorescence staining with F4/80 (green). Scale bar: 200 µm. The area and the F4/80 fluorescence intensity were quantified with ImageJ FIJI software and represented in the bar graph expressed as units relative to WT CNV+ vehicle control. Bars denote the mean ± SD from triplicate experiments, n = 4 mice/group. ns: non-significant.

Article Snippet: The following antibodies were used: rabbit polyclonal anti-p75 NTR (PRB-602C Covance, now Biolegend), rabbit polyclonal anti-GFAP (1/1000; Dako, Carpinteria, CA, USA), mouse monoclonal anti-β tubulin (1/5000; Sigma), mouse monoclonal anti-β-actin (1/2000; ab8226, Abcam).

Techniques: Immunofluorescence, Staining, Software, Control, Injection, Flow Cytometry, Fluorescence

Journal: BioMed Research International

Article Title: Imbalance of the Nerve Growth Factor and Its Precursor as a Potential Biomarker for Diabetic Retinopathy

doi: 10.1155/2015/571456

Figure Lengend Snippet: Shedding of p75 NTR is consistent in vitreous and serum. (a) Representative bands show p75 NTR expression in vitreous and serum for diabetic compared to nondiabetic control groups. The full length p75 NTR (75 kD) and receptor ectodomain (50 kD) had similar levels of expression in control and diabetic (DB) groups of both vitreous and serum. The possible proteolytic C terminal fragment (CTF) and intracellular domain (ICD) appeared at 27 kD and 22 kD. Differences in expression patterns between vitreous and serum as well as between diabetic and control groups were evident for both CTF and ICD. (b) In vitreous, 27 kD p75 NTR receptor fragment was significantly increased in diabetic (1.65-fold ± 0.23) compared to nondiabetic control group ( N = 4–11, * P < 0.05). (c) In serum, a significant increase in 22 kD p75 NTR occurred in diabetic samples (1.85-fold ± 0.30) compared to nondiabetic controls ( N = 6–10, * P < 0.05).

Article Snippet: The following antibodies were used for immunoblotting: rabbit polyclonal anti-NGF and anti-proNGF (Alomone Labs, Israel) and rabbit polyclonal p75 NTR (1 : 5000), a kind gift from Dr. Bruce Carter, Vanderbilt University, Nashville, TN.

Techniques: Expressing

Journal: BioMed Research International

Article Title: Imbalance of the Nerve Growth Factor and Its Precursor as a Potential Biomarker for Diabetic Retinopathy

doi: 10.1155/2015/571456

Figure Lengend Snippet: Expression of p75 NTR receptor is consistent in vitreous and serum. Results are shown for p75 NTR expression in vitreous and serum of diabetic (DB) and control participants normalized to Ponceau S and respective controls. Full length p75 NTR receptor (75 kD) was not significantly different in diabetic sample compared to control groups in either (a) vitreous ( N = 4–11) or (b) serum ( N = 6–9). The p75 NTR ectodomain (50 kD) was also not significantly different in diabetic compared to control groups in (c) vitreous ( N = 4–11) or (d) serum ( N = 6–9).

Article Snippet: The following antibodies were used for immunoblotting: rabbit polyclonal anti-NGF and anti-proNGF (Alomone Labs, Israel) and rabbit polyclonal p75 NTR (1 : 5000), a kind gift from Dr. Bruce Carter, Vanderbilt University, Nashville, TN.

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Clostridium butyricum RH2 Alleviates Chronic Foot Shock Stress-Induced Behavioral Deficits in Rats via PAI-1

doi: 10.3389/fphar.2022.845221

Figure Lengend Snippet: Effects of C. butyricum RH2 on the expression of proBDNF, P75 NTR , BDNF and TrkB in the hippocampus of CFSS rats. (A–C) proBDNF and P75 NTR were upregulated after CFSS but the increase was partially reversed by C. butyricum RH2. (D–F) BDNF and TrkB were downregulated after CFSS but the decrease was partially reversed by C. butyricum RH2. * p < 0.05, ** p < 0.01 versus sham group; # p < 0.05, ## p < 0.01 versus stress group; ^ p < 0.05, ^^ p < 0.01 versus stress + C. butyricum group.

Article Snippet: The membrane was then blocked with 5% milk in PBS-Tween 20 for 1 h, and incubated overnight at 4°C with mouse anti-GFAP (3670s, Cell Signaling Technology, United States; 1:1,000), rabbit anti-BDNF (ab108319, Abcam, United Kingdom; 1:1,000), rabbit anti-TrkB (ab33655, Abcam, United Kingdom; 1:1,000), rabbit anti-p75 NTR (ab52987, Abcam, United Kingdom; 1:1,000), anti-Pro-BDNF (sc-65513, SANTA CRUZ, USA; 1:200), mouse anti-β-Actin (AF0003, Beyotime, China; 1:1,000) antibodies, rabbit anti-PAI-1 (ab66705, Abcam, United Kingdom; 1:1,000) and rabbit anti-tPA (10147-1-AP, Proteintech, China, 1:1,000), respectively.

Techniques: Expressing