Review

cells r d systems af1157 goat polyclonal (R&D Systems)

R&D Systems is a verified supplier

R&D Systems manufactures this product

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

R&D Systems cells r d systems af1157 goat polyclonal
Cells R D Systems Af1157 Goat Polyclonal, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 43 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/goat+anti+mouse+ngfr+polyclonal+antibody/Mouse+NGFR%2FTNFRSF16+Antibody/pm39695302-438-71-72
Average 94 stars, based on 43 article reviews

cells r d systems af1157 goat polyclonal - by Bioz Stars, 2026-10

94/100 stars

Images

other:

Article Title: The aryl hydrocarbon receptor promotes inflammation-induced dedifferentiation and systemic metastatic spread of melanoma cells.
Article Snippet: Used antibodies were as follows: sheep antimouse AHR polyclonal antibody (R&D, Minneapolis, Minnesota, USA , AF6697), sheep anti-human AHR polyclonal antibody (R&D, AF6185), mouse β-Actin monoclonal antibody (Santa Cruz, Dallas, Texas, USA, sc-47778), goat anti-mouse NGFR polyclonal antibody (R&D, AF1157) goat antimouse/human gp100 polyclonal antibody (Abcam, Cambridge, UK, ab52058), rabbit antimouse/human/rat TYRP1 polyclonal antibody (Novus, Centennial, Colorado, USA, NBP1-88370), rabbit anti-mouse/human TRP2 polyclonal antibody (Abcam, ab74073), goat anti-rabbit IgG HRP-linked Antibody (Cell Signaling, #7074S), mouse anti-goat IgG HRP linked antibody (Santa Cruz, sc-2354) and donkey anti-sheep IgG HRP-linked antibody (R&D, HAF016).

Immunohistochemistry:

Article Title: A Preclinical Model of Malignant Peripheral Nerve Sheath Tumor-like Melanoma Is Characterized by Infiltrating Mast Cells
Article Snippet: Immunohistochemistry was performed with rabbit antimouse gp100 polyclonal antibody (Novus Biologicals; NBP169571) and goat anti-mouse NGFR polyclonal antibody (R&D Systems; BAF1157), followed by enzyme-conjugated secondary antibodies and the LSAB-2 color development system (DAKO).

Article Title: Melanomas resist T-cell therapy through inflammation-induced reversible dedifferentiation.
Article Snippet: Adoptive cell transfer therapies (ACTs) with cytotoxic T cells that target melanocytic antigens can achieve remissions in patients with metastatic melanomas, but tumours frequently relapse.. Hypotheses explaining the acquired resistance to ACTs include the selection of antigen-deficient tumour cell variants and the induction of T-cell tolerance.. However, the lack of appropriate experimental melanoma models has so far impeded clear insights into the underlying mechanisms.

Article Title: A Preclinical Model of Malignant Peripheral Nerve Sheath Tumor-like Melanoma Is Characterized by Infiltrating Mast Cells
Article Snippet: Human melanomas exhibit considerable genetic, pathological, and microenvironmental heterogeneity.. Genetically engineered mice have successfully been used to model the genomic aberrations contributing to melanoma pathogenesis, but their ability to recapitulate the phenotypic variability of human disease and the complex interactions with the immune system have not been addressed.. Here we report the unexpected finding that immune-cell poor pigmented and immunecell rich amelanotic melanomas developed simultaneously in Cdk4R24C mutant mice upon melanocyte-specific conditional activation of oncogenic BrafV600E and a single application of the carcinogen DMBA.

Image Search Results

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.

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

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.

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.

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

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.

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.

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

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.

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.

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

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.

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.

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

a) Schematic diagram represents CCI conducted on the left somatosensory cortex of mice at P90. b) A coronal section of mice brain stained with DAPI represents the region of implemented CCI. c) Immunostaining assay demonstrates BACE1 (green) and p75 (red) in both CCI and sham groups 72 hours after CCI. “Merge” is the combination of DAPI (blue), BACE1, and p75. White arrows indicate the cells, expressing both BACE1, and p75 within 300 μm from the injury site. The white dashes on the green channel indicate CCI region. n=6 mice, scale bar: 200 μm.

Journal: Molecular and cellular neurosciences

Article Title: Regulation of BACE1 expression after injury is linked to the p75 neurotrophin receptor

doi: 10.1016/j.mcn.2019.103395

Figure Lengend Snippet: a) Schematic diagram represents CCI conducted on the left somatosensory cortex of mice at P90. b) A coronal section of mice brain stained with DAPI represents the region of implemented CCI. c) Immunostaining assay demonstrates BACE1 (green) and p75 (red) in both CCI and sham groups 72 hours after CCI. “Merge” is the combination of DAPI (blue), BACE1, and p75. White arrows indicate the cells, expressing both BACE1, and p75 within 300 μm from the injury site. The white dashes on the green channel indicate CCI region. n=6 mice, scale bar: 200 μm.

Article Snippet: Goat polyclonal anti- p75 (Cat. No. AF1157, R&D Systems, Minneapolis, MN, USA), rabbit ranti-p75ICD (9991) ( Zampieri et al., 2005 ), anti-p75ECD antiserum (9651) ( Huber and Chao, 1995 ), rabbit anti-Fas receptor (M20, Cat. No. sc-716, Santa Cruz, Dallas, TX, USA), rabbit monoclonal anti- BACE1 (D10E5, Cat. No. 5606, Cell Signaling Technology, Danvers, MA, USA), rabbit SAPK/JNK (Cat No. 9252, Cell Signaling Technology), rabbit phospho-SAPK/JNK (Thr183/Tyr185) (Cat No. 9251, Cell Signaling Technology), mouse anti-glial fibrillary acidic protein (GFAP) (Cat. No. 556330, Biosciences, Bedford, MA, USA), chicken anti-NeuN (Cat No. ABN91, Millipore, Burlington, MA, USA), mouse anti-APP (22C11, Cat. No. MAB384) (Millipore, Burlington, MA, USA), anti-APP (C1/6.1 and m3.2) antibodies ( Choi et al., 2009 ), and mouse monoclonal anti- β- actin (AC-74, Cat. No. A2228, Sigma- Aldrich, St Louis, MO, USA) were used as primary antibodies in the present study.

Techniques: Staining, Immunostaining, Expressing

a, b) p75 and BACE1 mRNA levels were measured in SY5Y cells transfected with 1 μg mock or p75 using rt-qPCR. The GAPDH mRNA was measured as a loading control (n = 3 individual experiments, student’s t-test, mean ± SEM, *p<0.05, **p<0.01). c) A schematic diagram shows a rat full-length BACE1 promoter (1.54 kb) in pGL3 vector (pGL3-BACE1) used for the luciferase assay in this study. BACE1 promoter activity was measured in SY5Y cells transfected with: d) 1 μg mock, p75, or Fas receptor, e) 0, 0.25, 0.5, 1 μg p75, f) 1 μg mock or p75 plasmids and in the presence of 10 ng/ml BDNF, NGF or proNGF for 24 hours, g) p75CTF and p75ECD. Western blot analysis was conducted to demonstrate transfection efficiency of constructs. (n = 3 individual experiments, One-way ANOVA with Tukey test, mean ± SEM, *p<0.05, **p<0.01, RLU: relative light units, control: no treatment, ns: not significant).

Journal: Molecular and cellular neurosciences

Article Title: Regulation of BACE1 expression after injury is linked to the p75 neurotrophin receptor

doi: 10.1016/j.mcn.2019.103395

Figure Lengend Snippet: a, b) p75 and BACE1 mRNA levels were measured in SY5Y cells transfected with 1 μg mock or p75 using rt-qPCR. The GAPDH mRNA was measured as a loading control (n = 3 individual experiments, student’s t-test, mean ± SEM, *p<0.05, **p<0.01). c) A schematic diagram shows a rat full-length BACE1 promoter (1.54 kb) in pGL3 vector (pGL3-BACE1) used for the luciferase assay in this study. BACE1 promoter activity was measured in SY5Y cells transfected with: d) 1 μg mock, p75, or Fas receptor, e) 0, 0.25, 0.5, 1 μg p75, f) 1 μg mock or p75 plasmids and in the presence of 10 ng/ml BDNF, NGF or proNGF for 24 hours, g) p75CTF and p75ECD. Western blot analysis was conducted to demonstrate transfection efficiency of constructs. (n = 3 individual experiments, One-way ANOVA with Tukey test, mean ± SEM, *p<0.05, **p<0.01, RLU: relative light units, control: no treatment, ns: not significant).

Techniques: Transfection, Quantitative RT-PCR, Control, Plasmid Preparation, Luciferase, Activity Assay, Western Blot, Construct

BACE1 and p75 expression levels after 24, 48, 72, and 96 hours following CCI in the mouse brain BACE1 and p75 expression were quantified by immunostaining assay within 24, 48, 72, and 96 hours following CCI.

Journal: Molecular and cellular neurosciences

Article Title: Regulation of BACE1 expression after injury is linked to the p75 neurotrophin receptor

doi: 10.1016/j.mcn.2019.103395

Figure Lengend Snippet: BACE1 and p75 expression levels after 24, 48, 72, and 96 hours following CCI in the mouse brain BACE1 and p75 expression were quantified by immunostaining assay within 24, 48, 72, and 96 hours following CCI.

Techniques: Expressing, Immunostaining

A coronal section of basal forebrain in mice at P90 was stained for DAPI (blue), BACE1 (green) and p75 (red). “Merge” shows the three channels combined. BF: basal forebrain, n=6 mice, scale bar: 200 μm.

Journal: Molecular and cellular neurosciences

Article Title: Regulation of BACE1 expression after injury is linked to the p75 neurotrophin receptor

doi: 10.1016/j.mcn.2019.103395

Figure Lengend Snippet: A coronal section of basal forebrain in mice at P90 was stained for DAPI (blue), BACE1 (green) and p75 (red). “Merge” shows the three channels combined. BF: basal forebrain, n=6 mice, scale bar: 200 μm.

Techniques: Staining

a) The immunofluorescence staining for BACE1 (green), p75 (red), and NeuN (+) (magenta) was performed in both sham and CCI groups within 72 hours following CCI experiment. “Merge” is the combination of three channels. The blue signal in “Merge” is DAPI. White arrows indicate NeuN cells. b) The graph represents the percentage of NeuN (+) cells expressing both BACE1 and p75 within 300 μm from the injury site. (n = 6 mice, student’s t-test, mean ± SEM, **p<0.01, scale bar: 50 μm)

Journal: Molecular and cellular neurosciences

Article Title: Regulation of BACE1 expression after injury is linked to the p75 neurotrophin receptor

doi: 10.1016/j.mcn.2019.103395

Figure Lengend Snippet: a) The immunofluorescence staining for BACE1 (green), p75 (red), and NeuN (+) (magenta) was performed in both sham and CCI groups within 72 hours following CCI experiment. “Merge” is the combination of three channels. The blue signal in “Merge” is DAPI. White arrows indicate NeuN cells. b) The graph represents the percentage of NeuN (+) cells expressing both BACE1 and p75 within 300 μm from the injury site. (n = 6 mice, student’s t-test, mean ± SEM, **p<0.01, scale bar: 50 μm)

Techniques: Immunofluorescence, Staining, Expressing

a) The immunofluorescence staining for BACE1 (green), p75 (red), and GFAP (+) (magenta) was performed in both sham and CCI groups within 72 hours following CCI experiment. “Merge” is the combination of three channels. The blue signal in “Merge” is DAPI. White arrows indicate GFAP cells. b) The graph demonstrates the percentage of GFAP (+) cells expressing, both BACE1 and p75 within 300 μm from the injury site. (n = 6 mice, student’s t-test, mean ± SEM, ns: not significant, scale bar: 50 μm)

Journal: Molecular and cellular neurosciences

Article Title: Regulation of BACE1 expression after injury is linked to the p75 neurotrophin receptor

doi: 10.1016/j.mcn.2019.103395

Figure Lengend Snippet: a) The immunofluorescence staining for BACE1 (green), p75 (red), and GFAP (+) (magenta) was performed in both sham and CCI groups within 72 hours following CCI experiment. “Merge” is the combination of three channels. The blue signal in “Merge” is DAPI. White arrows indicate GFAP cells. b) The graph demonstrates the percentage of GFAP (+) cells expressing, both BACE1 and p75 within 300 μm from the injury site. (n = 6 mice, student’s t-test, mean ± SEM, ns: not significant, scale bar: 50 μm)

Techniques: Immunofluorescence, Staining, Expressing

A schematic diagram demonstrates a focal injury induced by a two-photon laser beam through a cranial window over the motor cortex of the mouse. b) The beam was placed at the desired position for approximately 1–3 seconds to create a small injury site as indicated by a bright autofluorescence sphere (~15 μm in diameter). c) After 72 hours of laser lesion, the immunofluorescence staining for BACE1 (green), p75 (red), GFAP (magenta), and DAPI (blue) was performed in ipsilateral and contralateral sites of the p75−/− and p75+/+ mice. The contralateral was not subjected to the laser injury experiment and considered as a control for ipsilateral. d) The fluorescence intensity of p75 and BACE1 within 200 μm from the lesion was quantified and plotted. (n = 6 mice, student’s t-test, mean ± SEM, **p<0.01, ns: not significant, scale bar: 50 μm).

Journal: Molecular and cellular neurosciences

Article Title: Regulation of BACE1 expression after injury is linked to the p75 neurotrophin receptor

doi: 10.1016/j.mcn.2019.103395

Figure Lengend Snippet: A schematic diagram demonstrates a focal injury induced by a two-photon laser beam through a cranial window over the motor cortex of the mouse. b) The beam was placed at the desired position for approximately 1–3 seconds to create a small injury site as indicated by a bright autofluorescence sphere (~15 μm in diameter). c) After 72 hours of laser lesion, the immunofluorescence staining for BACE1 (green), p75 (red), GFAP (magenta), and DAPI (blue) was performed in ipsilateral and contralateral sites of the p75−/− and p75+/+ mice. The contralateral was not subjected to the laser injury experiment and considered as a control for ipsilateral. d) The fluorescence intensity of p75 and BACE1 within 200 μm from the lesion was quantified and plotted. (n = 6 mice, student’s t-test, mean ± SEM, **p<0.01, ns: not significant, scale bar: 50 μm).

Techniques: Immunofluorescence, Staining, Control, Fluorescence

Western blot analysis was conducted to detect BACE1 in SY5Y (a) and HEK-293 (b) cells transfected with 1μg, either mock vector or p75 constructs. c) The graph represents the quantification of (a) and (b). The cultured cortical neurons (DIV-III) isolated from E18 p75−/− and p75+/+ mice were subjected to immunostaining (d and e) and Western blotting (f and g) assays for BACE1 analysis. (n = 3 individual experiments, student’s t-test, mean ± SEM, *p<0.05, scale bar: 50 μm). β-actin in Western blot was used as a loading control.

Journal: Molecular and cellular neurosciences

Article Title: Regulation of BACE1 expression after injury is linked to the p75 neurotrophin receptor

doi: 10.1016/j.mcn.2019.103395

Figure Lengend Snippet: Western blot analysis was conducted to detect BACE1 in SY5Y (a) and HEK-293 (b) cells transfected with 1μg, either mock vector or p75 constructs. c) The graph represents the quantification of (a) and (b). The cultured cortical neurons (DIV-III) isolated from E18 p75−/− and p75+/+ mice were subjected to immunostaining (d and e) and Western blotting (f and g) assays for BACE1 analysis. (n = 3 individual experiments, student’s t-test, mean ± SEM, *p<0.05, scale bar: 50 μm). β-actin in Western blot was used as a loading control.

Techniques: Western Blot, Transfection, Plasmid Preparation, Construct, Cell Culture, Isolation, Immunostaining, Control

a) SY5Y-APP cells were transfected with 0.5 and 1 μg p75 or mock constructs and then subjected to Western blot analysis to detect p75 and BACE1 levels. Additionally, the full length and CTFs of APP were detected by C1/6.1 antibody. To detect soluble fragments of APP, the conditioned media was subjected to immunoprecipitation (IP) using m3.2 antibody. The IP and flow-through samples were then subjected to Western blot using anti-APP (22C11). The blotting signals obtained from IP and flow-through, represent the level of sAPPα and sAPPβ, respectively (Choi et al., 2009). b-d) Graphs demonstrate the quantification of (a). (n = 3 individual experiments, One-way ANOVA with Tukey test, mean ± SEM, *p<0.05, **p<0.01). e) Cortical neuronal isolated from E18 p75−/− and p75+/+ mice at DIV-III were lysed and subjected to Western blot to detect BACE1, fl.APP, and sAPPβ levels. f, g) The quantification of (e) (n = 3 individual experiments, student’s t-test, mean ± SEM, *p<0.05). fl.APP and β-actin were used as loading controls.

Journal: Molecular and cellular neurosciences

Article Title: Regulation of BACE1 expression after injury is linked to the p75 neurotrophin receptor

doi: 10.1016/j.mcn.2019.103395

Figure Lengend Snippet: a) SY5Y-APP cells were transfected with 0.5 and 1 μg p75 or mock constructs and then subjected to Western blot analysis to detect p75 and BACE1 levels. Additionally, the full length and CTFs of APP were detected by C1/6.1 antibody. To detect soluble fragments of APP, the conditioned media was subjected to immunoprecipitation (IP) using m3.2 antibody. The IP and flow-through samples were then subjected to Western blot using anti-APP (22C11). The blotting signals obtained from IP and flow-through, represent the level of sAPPα and sAPPβ, respectively (Choi et al., 2009). b-d) Graphs demonstrate the quantification of (a). (n = 3 individual experiments, One-way ANOVA with Tukey test, mean ± SEM, *p<0.05, **p<0.01). e) Cortical neuronal isolated from E18 p75−/− and p75+/+ mice at DIV-III were lysed and subjected to Western blot to detect BACE1, fl.APP, and sAPPβ levels. f, g) The quantification of (e) (n = 3 individual experiments, student’s t-test, mean ± SEM, *p<0.05). fl.APP and β-actin were used as loading controls.

Techniques: Transfection, Construct, Western Blot, Immunoprecipitation, Isolation

a) A schematic diagram represents the full-length and mutant (ΔNF-κB) rat BACE1 promoter constructs. b) The promoter activity of full-length or ΔNF-κB BACE1 promoter was measured in the SY5Y cells transfected with either mock or p75 constructs. c-e) SY5Y cells were transfected with mock and p75 constructs for 24 hours and then treated with 10 μg/mL SP600125, for an additional 3 hours. The lysate was subjected to the luciferase assay (c) and Western blot analysis (d,e) to measure BACE1 promoter activity and expression levels, respectively. f,g) The level of pJNK in SY5Y cells transfected with 0, 0.5, and 1 μg of p75 plasmid was measured by Western blot analysis. SY5Y cells were exposed with U.V light for 3 hours to activate JNK and the cell lysate was used as a positive control for anti-pJNK antibody. (n = 3 individual experiments, One-way ANOVA with Tukey test, mean ± SEM, *p<0.05, **p<0.01, RLU: relative light units, cont: no treatment). h) Western blot was performed on cultured cortical neurons (DIV-III) isolated from E18 p75−/− and p75+/+ mice to measure pJNK, JNK (total), and BACE1 levels. i,j) Graphs are the quantification of (h). β-actin and JNK were considered as loading controls. (n = 3 individual experiments, student’s t-test, mean ± SEM, *p<0.05, RLU: relative light units)

Journal: Molecular and cellular neurosciences

Article Title: Regulation of BACE1 expression after injury is linked to the p75 neurotrophin receptor

doi: 10.1016/j.mcn.2019.103395

Figure Lengend Snippet: a) A schematic diagram represents the full-length and mutant (ΔNF-κB) rat BACE1 promoter constructs. b) The promoter activity of full-length or ΔNF-κB BACE1 promoter was measured in the SY5Y cells transfected with either mock or p75 constructs. c-e) SY5Y cells were transfected with mock and p75 constructs for 24 hours and then treated with 10 μg/mL SP600125, for an additional 3 hours. The lysate was subjected to the luciferase assay (c) and Western blot analysis (d,e) to measure BACE1 promoter activity and expression levels, respectively. f,g) The level of pJNK in SY5Y cells transfected with 0, 0.5, and 1 μg of p75 plasmid was measured by Western blot analysis. SY5Y cells were exposed with U.V light for 3 hours to activate JNK and the cell lysate was used as a positive control for anti-pJNK antibody. (n = 3 individual experiments, One-way ANOVA with Tukey test, mean ± SEM, *p<0.05, **p<0.01, RLU: relative light units, cont: no treatment). h) Western blot was performed on cultured cortical neurons (DIV-III) isolated from E18 p75−/− and p75+/+ mice to measure pJNK, JNK (total), and BACE1 levels. i,j) Graphs are the quantification of (h). β-actin and JNK were considered as loading controls. (n = 3 individual experiments, student’s t-test, mean ± SEM, *p<0.05, RLU: relative light units)

Techniques: Mutagenesis, Construct, Activity Assay, Transfection, Luciferase, Western Blot, Expressing, Plasmid Preparation, Positive Control, Cell Culture, Isolation

Review

cells r d systems af1157 goat polyclonal (R&D Systems)

Structured Review

Images

Related Articles

other:

Immunohistochemistry:

Similar Products

Image Search Results