p75ntr  (Alomone Labs)


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    Structured Review

    Alomone Labs p75ntr
    Necdin is coexpressed with TrkA or <t>p75NTR</t> in developing DRGs. A , B , Double immunostaining for necdin and TrkA or p75NTR. Frozen DRG sections of E13.5 mouse embryo were double immunostained for necdin with antibody GN1 (red) and TrkA (or p75NTR) (green), and two images were merged (yellow). Scale bars: A , 50 μm; B , 10 μm. C , Immunoaffinity assay for endogenous necdin-interacting proteins. Tissue lysate (1 mg) of the DRG and spinal cord from E13.5 embryos was applied to the immunoaffinity columns with anti-necdin IgG (NC243) (αNecdin IgG) and control preimmune IgG (Preimmune IgG). The eluates were analyzed by Western blotting for endogenous complexes of necdin with TrkA, p75NTR, and APP. Lysate, Tissue lysate (30 μg).
    P75ntr, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p75ntr/product/Alomone Labs
    Average 94 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    p75ntr - by Bioz Stars, 2022-08
    94/100 stars

    Images

    1) Product Images from "Disruption of the Paternal Necdin Gene Diminishes TrkA Signaling for Sensory Neuron Survival"

    Article Title: Disruption of the Paternal Necdin Gene Diminishes TrkA Signaling for Sensory Neuron Survival

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.2083-05.2005

    Necdin is coexpressed with TrkA or p75NTR in developing DRGs. A , B , Double immunostaining for necdin and TrkA or p75NTR. Frozen DRG sections of E13.5 mouse embryo were double immunostained for necdin with antibody GN1 (red) and TrkA (or p75NTR) (green), and two images were merged (yellow). Scale bars: A , 50 μm; B , 10 μm. C , Immunoaffinity assay for endogenous necdin-interacting proteins. Tissue lysate (1 mg) of the DRG and spinal cord from E13.5 embryos was applied to the immunoaffinity columns with anti-necdin IgG (NC243) (αNecdin IgG) and control preimmune IgG (Preimmune IgG). The eluates were analyzed by Western blotting for endogenous complexes of necdin with TrkA, p75NTR, and APP. Lysate, Tissue lysate (30 μg).
    Figure Legend Snippet: Necdin is coexpressed with TrkA or p75NTR in developing DRGs. A , B , Double immunostaining for necdin and TrkA or p75NTR. Frozen DRG sections of E13.5 mouse embryo were double immunostained for necdin with antibody GN1 (red) and TrkA (or p75NTR) (green), and two images were merged (yellow). Scale bars: A , 50 μm; B , 10 μm. C , Immunoaffinity assay for endogenous necdin-interacting proteins. Tissue lysate (1 mg) of the DRG and spinal cord from E13.5 embryos was applied to the immunoaffinity columns with anti-necdin IgG (NC243) (αNecdin IgG) and control preimmune IgG (Preimmune IgG). The eluates were analyzed by Western blotting for endogenous complexes of necdin with TrkA, p75NTR, and APP. Lysate, Tissue lysate (30 μg).

    Techniques Used: Double Immunostaining, Western Blot

    Necdin interacts with both TrkA and p75NTR. A , B , Immunoprecipitation assay for interactions of necdin with TrkA and p75NTR. Lysates of HEK293A cells transfected with expression vectors for FLAG-TrkA, FLAG-p75NTR, FLAG-APP, and necdin were immunoprecipitated (IP) with anti-FLAG antibody M2 (αFLAG) and immunoblotted (IB) with anti-necdin antibody NC243 (αNecdin) ( A , top panel). Conversely, the lysates were immunoprecipitated with antibody NC243 and immunoblotted with antibody M2 ( B , top panel). Expressed proteins in cell lysates are shown in the bottom panels. C , Necdin-enhanced association between TrkA and p75NTR. Lysates of cells transfected with expression vectors for HA-p75NTR (4 μg), FLAG-TrkA (4.5 μg), and necdin (0.1 and 0.5 μg) were immunoprecipitated with antibody M2 and immunoblotted with anti-p75NTR antibody (top panel). D , Interactions of necdin with TrkA mutants. Lysates of cDNA-transfected cells expressing necdin, FLAG-tagged wild-type (WT) TrkA, and the mutants (K547A, Y499A, Y794A) were immunoprecipitated as in A . Phosphorylated Tyr499 in rat TrkA was analyzed using the antibody against phospho-human TrkA (Tyr490) (αpTrkA) ( D , second panel from the top).
    Figure Legend Snippet: Necdin interacts with both TrkA and p75NTR. A , B , Immunoprecipitation assay for interactions of necdin with TrkA and p75NTR. Lysates of HEK293A cells transfected with expression vectors for FLAG-TrkA, FLAG-p75NTR, FLAG-APP, and necdin were immunoprecipitated (IP) with anti-FLAG antibody M2 (αFLAG) and immunoblotted (IB) with anti-necdin antibody NC243 (αNecdin) ( A , top panel). Conversely, the lysates were immunoprecipitated with antibody NC243 and immunoblotted with antibody M2 ( B , top panel). Expressed proteins in cell lysates are shown in the bottom panels. C , Necdin-enhanced association between TrkA and p75NTR. Lysates of cells transfected with expression vectors for HA-p75NTR (4 μg), FLAG-TrkA (4.5 μg), and necdin (0.1 and 0.5 μg) were immunoprecipitated with antibody M2 and immunoblotted with anti-p75NTR antibody (top panel). D , Interactions of necdin with TrkA mutants. Lysates of cDNA-transfected cells expressing necdin, FLAG-tagged wild-type (WT) TrkA, and the mutants (K547A, Y499A, Y794A) were immunoprecipitated as in A . Phosphorylated Tyr499 in rat TrkA was analyzed using the antibody against phospho-human TrkA (Tyr490) (αpTrkA) ( D , second panel from the top).

    Techniques Used: Immunoprecipitation, Transfection, Expressing

    Necdin enhances NGF/TrkA signaling in PC12 cells. A , Coimmunoprecipitation assay for the association between TrkA and p75NTR. PC12 cells infected with recombinant adenoviruses expressing necdin (Ad-Necdin) and β-galactosidase (Ad-LacZ) were treated with NGF (50 ng/ml) for 38 h. Cell lysates (400 μg) were immunoprecipitated (IP) with mouse anti-p75NTR antibody MC192 (αp75NTRm) and immunoblotted (IB) with anti-TrkA antibody (top panel). Expressed proteins in cell lysates are shown in the bottom panels. B , Neurite outgrowth assay. Ad-Necdin- and Ad-LacZ-infected PC12 cells were treated with NGF for 38 h, and cells bearing extended neurites were counted (mean ± SEM; n = 4; p
    Figure Legend Snippet: Necdin enhances NGF/TrkA signaling in PC12 cells. A , Coimmunoprecipitation assay for the association between TrkA and p75NTR. PC12 cells infected with recombinant adenoviruses expressing necdin (Ad-Necdin) and β-galactosidase (Ad-LacZ) were treated with NGF (50 ng/ml) for 38 h. Cell lysates (400 μg) were immunoprecipitated (IP) with mouse anti-p75NTR antibody MC192 (αp75NTRm) and immunoblotted (IB) with anti-TrkA antibody (top panel). Expressed proteins in cell lysates are shown in the bottom panels. B , Neurite outgrowth assay. Ad-Necdin- and Ad-LacZ-infected PC12 cells were treated with NGF for 38 h, and cells bearing extended neurites were counted (mean ± SEM; n = 4; p

    Techniques Used: Co-Immunoprecipitation Assay, Infection, Recombinant, Expressing, Immunoprecipitation, Neurite Outgrowth Assay

    Necdin expression is absent in DRGs of mutant mice lacking the paternal necdin gene. A , The restriction map of mouse Ndn locus. A 7 kb Ndn sequence containing coding (black) and 3′,5′ noncoding (hatched) regions was subcloned into the targeting vector. The 1.5 kb expression unit containing the pgk promoter with Neo r gene (Pgk1/neo r ), polyadenylation signal [Poly(A) signal], was inserted to disrupt the Ndn coding sequence. The vector was introduced to TT2 cells for homologous recombination as described in Materials and Methods. B , Absence of necdin expression in the brain and spinal cord of mice deficient in the paternal necdin gene. Frozen sections of the hypothalamus and spinal cord from E13.5 wild-type (Ndn +m/+p ) and necdin-deficient (Ndn +m/-p ) littermates were immunostained for necdin. HT, Hypothalamus; SC, spinal cord; DBB, diagonal band of Broca; ML, mantle layer. Scale bars: HT, 50 μm; SC, 100 μm. C , Expression patterns of TrkA and p75NTR in necdin-deficient DRGs. Frozen sections of cervical DRGs from E12.5 wild-type (Ndn +m/+p ) and necdin-deficient (Ndn +m/-p ) littermates were immunostained for necdin, TrkA, and p75NTR. Scale bar, 50 μm. D , E , Western blot analysis of necdin, TrkA, p75NTR, and phosphorylated MAPK. Equal amounts of DRG lysates from E12.5 embryos were analyzed by Western blotting. Each lane represents pooled DRGs from a single embryo. Molecular sizes in kilodaltons are shown to the right. E , Signal intensities of TrkA, p75NTR, and pMAPK shown in D were normalized to those of tubulin (TB) and MAPK (mean ± SEM; n = 5 for each protein). * p
    Figure Legend Snippet: Necdin expression is absent in DRGs of mutant mice lacking the paternal necdin gene. A , The restriction map of mouse Ndn locus. A 7 kb Ndn sequence containing coding (black) and 3′,5′ noncoding (hatched) regions was subcloned into the targeting vector. The 1.5 kb expression unit containing the pgk promoter with Neo r gene (Pgk1/neo r ), polyadenylation signal [Poly(A) signal], was inserted to disrupt the Ndn coding sequence. The vector was introduced to TT2 cells for homologous recombination as described in Materials and Methods. B , Absence of necdin expression in the brain and spinal cord of mice deficient in the paternal necdin gene. Frozen sections of the hypothalamus and spinal cord from E13.5 wild-type (Ndn +m/+p ) and necdin-deficient (Ndn +m/-p ) littermates were immunostained for necdin. HT, Hypothalamus; SC, spinal cord; DBB, diagonal band of Broca; ML, mantle layer. Scale bars: HT, 50 μm; SC, 100 μm. C , Expression patterns of TrkA and p75NTR in necdin-deficient DRGs. Frozen sections of cervical DRGs from E12.5 wild-type (Ndn +m/+p ) and necdin-deficient (Ndn +m/-p ) littermates were immunostained for necdin, TrkA, and p75NTR. Scale bar, 50 μm. D , E , Western blot analysis of necdin, TrkA, p75NTR, and phosphorylated MAPK. Equal amounts of DRG lysates from E12.5 embryos were analyzed by Western blotting. Each lane represents pooled DRGs from a single embryo. Molecular sizes in kilodaltons are shown to the right. E , Signal intensities of TrkA, p75NTR, and pMAPK shown in D were normalized to those of tubulin (TB) and MAPK (mean ± SEM; n = 5 for each protein). * p

    Techniques Used: Expressing, Mutagenesis, Mouse Assay, Sequencing, Plasmid Preparation, Homologous Recombination, Western Blot

    2) Product Images from "Differential Hippocampal Expression of BDNF Isoforms and Their Receptors Under Diverse Configurations of the Serotonergic System in a Mice Model of Increased Neuronal Survival"

    Article Title: Differential Hippocampal Expression of BDNF Isoforms and Their Receptors Under Diverse Configurations of the Serotonergic System in a Mice Model of Increased Neuronal Survival

    Journal: Frontiers in Cellular Neuroscience

    doi: 10.3389/fncel.2019.00384

    Effect of 8-OH-DPAT on the protein expression of molecules from the BDNF signaling pathway in the HC, determined by Western Blot. (A) mBDNF. (B) TrkB. (C) proBDNF. (D) p75 in hyposerotonergic (PCPA-treated) or control (vehicle-treated) mice receiving the 5-HT 1 A agonist 8-OH-DPAT (striped bars) or NaCl (white bars). (E) Representative membrane showing signal for p75 (75 kDa) and tubuline (52 kDa). 1: Control-NaCl. 2: Control-DPAT. 3: PCPA-NaCl. 4: PCPA-DPAT. Data are expressed as mean ± S.E.M., n = 6 per experimental group. ∗ p
    Figure Legend Snippet: Effect of 8-OH-DPAT on the protein expression of molecules from the BDNF signaling pathway in the HC, determined by Western Blot. (A) mBDNF. (B) TrkB. (C) proBDNF. (D) p75 in hyposerotonergic (PCPA-treated) or control (vehicle-treated) mice receiving the 5-HT 1 A agonist 8-OH-DPAT (striped bars) or NaCl (white bars). (E) Representative membrane showing signal for p75 (75 kDa) and tubuline (52 kDa). 1: Control-NaCl. 2: Control-DPAT. 3: PCPA-NaCl. 4: PCPA-DPAT. Data are expressed as mean ± S.E.M., n = 6 per experimental group. ∗ p

    Techniques Used: Expressing, Western Blot, Mouse Assay

    Protein expression of molecules from the BDNF signaling pathway determined by Western Blot and representative membranes. (A) mBDNF. (B) TrkB. (C) proBDNF. (D) p75 in hyposerotonergic (PCPA-treated) or control (vehicle-treated) mice for 4 weeks. (E–H) Representative membrane showing signal for TrkB, p75, pro-BDNF, and mBDNF in Control and PCPA-treatd mice. Data are expressed as mean ± S.E.M., n = 11 (control) and 12 (PCPA). ∗ p
    Figure Legend Snippet: Protein expression of molecules from the BDNF signaling pathway determined by Western Blot and representative membranes. (A) mBDNF. (B) TrkB. (C) proBDNF. (D) p75 in hyposerotonergic (PCPA-treated) or control (vehicle-treated) mice for 4 weeks. (E–H) Representative membrane showing signal for TrkB, p75, pro-BDNF, and mBDNF in Control and PCPA-treatd mice. Data are expressed as mean ± S.E.M., n = 11 (control) and 12 (PCPA). ∗ p

    Techniques Used: Expressing, Western Blot, Mouse Assay

    3) Product Images from "Influence of bacterial components on the developmental programming of enteric neurons, et al. Influence of bacterial components on the developmental programming of enteric neurons"

    Article Title: Influence of bacterial components on the developmental programming of enteric neurons, et al. Influence of bacterial components on the developmental programming of enteric neurons

    Journal: Physiological Reports

    doi: 10.14814/phy2.14611

    Characterization of ENCDC cultures. Flow cytometry of ENCDC cultures stained with antibodies against the immature neuronal marker, p75 NTR+ ‐FITC demonstrated 95.1% p75 NTR+ ‐positive, 97.3% viability in subculture 3, and 96.1% p75 NTR+ ‐positive, 98.2% viability in subculture 4 (a). Proportion of ENCDC cultures characterized for expression of p75NTR+, pH3, 5‐HT, nNOS, and TH. Scale bar = 20 µm (b). There was a significant increase in the proportion of serotonergic neurons and nitrergic neurons in subculture 5 (gray) compared to subculture 4 (black), but no significant difference in proliferating cells or dopaminergic neurons across the cultures. *p ≤ .05. Values are presented as mean ± SEM (c)
    Figure Legend Snippet: Characterization of ENCDC cultures. Flow cytometry of ENCDC cultures stained with antibodies against the immature neuronal marker, p75 NTR+ ‐FITC demonstrated 95.1% p75 NTR+ ‐positive, 97.3% viability in subculture 3, and 96.1% p75 NTR+ ‐positive, 98.2% viability in subculture 4 (a). Proportion of ENCDC cultures characterized for expression of p75NTR+, pH3, 5‐HT, nNOS, and TH. Scale bar = 20 µm (b). There was a significant increase in the proportion of serotonergic neurons and nitrergic neurons in subculture 5 (gray) compared to subculture 4 (black), but no significant difference in proliferating cells or dopaminergic neurons across the cultures. *p ≤ .05. Values are presented as mean ± SEM (c)

    Techniques Used: Flow Cytometry, Staining, Marker, Expressing

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    Alomone Labs anti p75ntr polyclonal antibody
    Process of differentiation of H-iris iPS cells to nervous system cells. Cells were pre-differentiated from ( a ) H-iris iPS cells to ( b ) neural stem/progenitor cells by adhesive culture. ( c ) Cells sorted with <t>p75NTR</t> were differentiated into neurons. By changing the medium condition, the nerve cells were differentiated into Recoverin-positive cells. ( d ) In contrast, after suspension culture, neurites were elongated by adhesive culture and differentiated into retinal ganglion cells. Bars: 100 μm.
    Anti P75ntr Polyclonal Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p75ntr polyclonal antibody/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti p75ntr polyclonal antibody - by Bioz Stars, 2022-08
    94/100 stars
      Buy from Supplier

    90
    Alomone Labs anti p75ntr
    Suggested model. In order to arrive at the cell body and subsequently the CNS, rabies virus hijacks a fast route using the <t>p75NTR</t> endosomal pathway. In a p75NTR dependent path, RABV manipulates the axonal transport machinery to migrate faster to the cell body. An alternative, slower path, may involve alternative RABV receptors.
    Anti P75ntr, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p75ntr/product/Alomone Labs
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti p75ntr - by Bioz Stars, 2022-08
    90/100 stars
      Buy from Supplier

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    Process of differentiation of H-iris iPS cells to nervous system cells. Cells were pre-differentiated from ( a ) H-iris iPS cells to ( b ) neural stem/progenitor cells by adhesive culture. ( c ) Cells sorted with p75NTR were differentiated into neurons. By changing the medium condition, the nerve cells were differentiated into Recoverin-positive cells. ( d ) In contrast, after suspension culture, neurites were elongated by adhesive culture and differentiated into retinal ganglion cells. Bars: 100 μm.

    Journal: Cells

    Article Title: Novel Technique for Retinal Nerve Cell Regeneration with Electrophysiological Functions Using Human Iris-Derived iPS Cells

    doi: 10.3390/cells10040743

    Figure Lengend Snippet: Process of differentiation of H-iris iPS cells to nervous system cells. Cells were pre-differentiated from ( a ) H-iris iPS cells to ( b ) neural stem/progenitor cells by adhesive culture. ( c ) Cells sorted with p75NTR were differentiated into neurons. By changing the medium condition, the nerve cells were differentiated into Recoverin-positive cells. ( d ) In contrast, after suspension culture, neurites were elongated by adhesive culture and differentiated into retinal ganglion cells. Bars: 100 μm.

    Article Snippet: Paraffin sections were prepared from the fixed human iris tissue in the usual manner and incubated with anti-p75NTR polyclonal antibody (1:200; Alomone Labs, Jerusalem, Israel) for 1 h at 37 °C.

    Techniques:

    Cells sorted with p75NTR can concentrate Recoverin-positive cells. ( a ) Before p75NTR sorting, cells with large cytoplasms ( arrowheads ) were mixed. ( b ) Fixed cells were analyzed for p75NTR and Nestin by FCM. Most of the cells sorted in the strong positive region of p75NTR were double-positive for ( c ) p75NTR and ( d ) Nestin. ( e , f ) The morphology of cells sorted by their strong positivity for ( e ) p75NTR or ( f ) their weak positivity or negativity for p75NTR. ( g ) Relative semi-quantitative analysis of Recoverin gene expression in cells selected by p75NTR, and pre/post-differentiated cells. ( h ) Fluorescent immunostaining of Recoverin in differentiated cells. Bar in panel ( a ): 50 μm, bars in panels ( c , d ): 20 μm, bars in panels ( e , f , h ): 100 μm.

    Journal: Cells

    Article Title: Novel Technique for Retinal Nerve Cell Regeneration with Electrophysiological Functions Using Human Iris-Derived iPS Cells

    doi: 10.3390/cells10040743

    Figure Lengend Snippet: Cells sorted with p75NTR can concentrate Recoverin-positive cells. ( a ) Before p75NTR sorting, cells with large cytoplasms ( arrowheads ) were mixed. ( b ) Fixed cells were analyzed for p75NTR and Nestin by FCM. Most of the cells sorted in the strong positive region of p75NTR were double-positive for ( c ) p75NTR and ( d ) Nestin. ( e , f ) The morphology of cells sorted by their strong positivity for ( e ) p75NTR or ( f ) their weak positivity or negativity for p75NTR. ( g ) Relative semi-quantitative analysis of Recoverin gene expression in cells selected by p75NTR, and pre/post-differentiated cells. ( h ) Fluorescent immunostaining of Recoverin in differentiated cells. Bar in panel ( a ): 50 μm, bars in panels ( c , d ): 20 μm, bars in panels ( e , f , h ): 100 μm.

    Article Snippet: Paraffin sections were prepared from the fixed human iris tissue in the usual manner and incubated with anti-p75NTR polyclonal antibody (1:200; Alomone Labs, Jerusalem, Israel) for 1 h at 37 °C.

    Techniques: Expressing, Immunostaining

    p75NTR-positive cells observed in human iris tissue and cultured cells: ( a ) HE-stained human iris tissue; * The lens side of the iris. ( b ) Differential interference contrast image of iris tissue. ( c ) Fluorescent immunostaining of p75NTR; arrowheads = p75NTR-positive cells. ( d ) Iris-derived cells were cultured for 10 days; the inset shows cells growing from around the pigmented cells on the third day of culture. ( e ) p75NTR-positive cells (region of Gate 1, G1) were isolated with a cell sorter; the blue line represents the histogram of negative cells. ( f ) Morphology of p75NTR-positive sorted cells. Bars in panels ( a – c ), 50 μm; bars in panel ( d )’s inset and panel ( f ), 100 μm; bar in panel ( d ), 200 μm.

    Journal: Cells

    Article Title: Novel Technique for Retinal Nerve Cell Regeneration with Electrophysiological Functions Using Human Iris-Derived iPS Cells

    doi: 10.3390/cells10040743

    Figure Lengend Snippet: p75NTR-positive cells observed in human iris tissue and cultured cells: ( a ) HE-stained human iris tissue; * The lens side of the iris. ( b ) Differential interference contrast image of iris tissue. ( c ) Fluorescent immunostaining of p75NTR; arrowheads = p75NTR-positive cells. ( d ) Iris-derived cells were cultured for 10 days; the inset shows cells growing from around the pigmented cells on the third day of culture. ( e ) p75NTR-positive cells (region of Gate 1, G1) were isolated with a cell sorter; the blue line represents the histogram of negative cells. ( f ) Morphology of p75NTR-positive sorted cells. Bars in panels ( a – c ), 50 μm; bars in panel ( d )’s inset and panel ( f ), 100 μm; bar in panel ( d ), 200 μm.

    Article Snippet: Paraffin sections were prepared from the fixed human iris tissue in the usual manner and incubated with anti-p75NTR polyclonal antibody (1:200; Alomone Labs, Jerusalem, Israel) for 1 h at 37 °C.

    Techniques: Cell Culture, Staining, Immunostaining, Derivative Assay, Isolation

    Necdin is coexpressed with TrkA or p75NTR in developing DRGs. A , B , Double immunostaining for necdin and TrkA or p75NTR. Frozen DRG sections of E13.5 mouse embryo were double immunostained for necdin with antibody GN1 (red) and TrkA (or p75NTR) (green), and two images were merged (yellow). Scale bars: A , 50 μm; B , 10 μm. C , Immunoaffinity assay for endogenous necdin-interacting proteins. Tissue lysate (1 mg) of the DRG and spinal cord from E13.5 embryos was applied to the immunoaffinity columns with anti-necdin IgG (NC243) (αNecdin IgG) and control preimmune IgG (Preimmune IgG). The eluates were analyzed by Western blotting for endogenous complexes of necdin with TrkA, p75NTR, and APP. Lysate, Tissue lysate (30 μg).

    Journal: The Journal of Neuroscience

    Article Title: Disruption of the Paternal Necdin Gene Diminishes TrkA Signaling for Sensory Neuron Survival

    doi: 10.1523/JNEUROSCI.2083-05.2005

    Figure Lengend Snippet: Necdin is coexpressed with TrkA or p75NTR in developing DRGs. A , B , Double immunostaining for necdin and TrkA or p75NTR. Frozen DRG sections of E13.5 mouse embryo were double immunostained for necdin with antibody GN1 (red) and TrkA (or p75NTR) (green), and two images were merged (yellow). Scale bars: A , 50 μm; B , 10 μm. C , Immunoaffinity assay for endogenous necdin-interacting proteins. Tissue lysate (1 mg) of the DRG and spinal cord from E13.5 embryos was applied to the immunoaffinity columns with anti-necdin IgG (NC243) (αNecdin IgG) and control preimmune IgG (Preimmune IgG). The eluates were analyzed by Western blotting for endogenous complexes of necdin with TrkA, p75NTR, and APP. Lysate, Tissue lysate (30 μg).

    Article Snippet: Antibodies used for coimmunoprecipitation assay are as follows: FLAG (M2; 1:50; Sigma), Necdin (NC243; 1:50), and p75NTR (MC192; 1:10; Alomone Labs, Jerusalem, Israel).

    Techniques: Double Immunostaining, Western Blot

    Necdin interacts with both TrkA and p75NTR. A , B , Immunoprecipitation assay for interactions of necdin with TrkA and p75NTR. Lysates of HEK293A cells transfected with expression vectors for FLAG-TrkA, FLAG-p75NTR, FLAG-APP, and necdin were immunoprecipitated (IP) with anti-FLAG antibody M2 (αFLAG) and immunoblotted (IB) with anti-necdin antibody NC243 (αNecdin) ( A , top panel). Conversely, the lysates were immunoprecipitated with antibody NC243 and immunoblotted with antibody M2 ( B , top panel). Expressed proteins in cell lysates are shown in the bottom panels. C , Necdin-enhanced association between TrkA and p75NTR. Lysates of cells transfected with expression vectors for HA-p75NTR (4 μg), FLAG-TrkA (4.5 μg), and necdin (0.1 and 0.5 μg) were immunoprecipitated with antibody M2 and immunoblotted with anti-p75NTR antibody (top panel). D , Interactions of necdin with TrkA mutants. Lysates of cDNA-transfected cells expressing necdin, FLAG-tagged wild-type (WT) TrkA, and the mutants (K547A, Y499A, Y794A) were immunoprecipitated as in A . Phosphorylated Tyr499 in rat TrkA was analyzed using the antibody against phospho-human TrkA (Tyr490) (αpTrkA) ( D , second panel from the top).

    Journal: The Journal of Neuroscience

    Article Title: Disruption of the Paternal Necdin Gene Diminishes TrkA Signaling for Sensory Neuron Survival

    doi: 10.1523/JNEUROSCI.2083-05.2005

    Figure Lengend Snippet: Necdin interacts with both TrkA and p75NTR. A , B , Immunoprecipitation assay for interactions of necdin with TrkA and p75NTR. Lysates of HEK293A cells transfected with expression vectors for FLAG-TrkA, FLAG-p75NTR, FLAG-APP, and necdin were immunoprecipitated (IP) with anti-FLAG antibody M2 (αFLAG) and immunoblotted (IB) with anti-necdin antibody NC243 (αNecdin) ( A , top panel). Conversely, the lysates were immunoprecipitated with antibody NC243 and immunoblotted with antibody M2 ( B , top panel). Expressed proteins in cell lysates are shown in the bottom panels. C , Necdin-enhanced association between TrkA and p75NTR. Lysates of cells transfected with expression vectors for HA-p75NTR (4 μg), FLAG-TrkA (4.5 μg), and necdin (0.1 and 0.5 μg) were immunoprecipitated with antibody M2 and immunoblotted with anti-p75NTR antibody (top panel). D , Interactions of necdin with TrkA mutants. Lysates of cDNA-transfected cells expressing necdin, FLAG-tagged wild-type (WT) TrkA, and the mutants (K547A, Y499A, Y794A) were immunoprecipitated as in A . Phosphorylated Tyr499 in rat TrkA was analyzed using the antibody against phospho-human TrkA (Tyr490) (αpTrkA) ( D , second panel from the top).

    Article Snippet: Antibodies used for coimmunoprecipitation assay are as follows: FLAG (M2; 1:50; Sigma), Necdin (NC243; 1:50), and p75NTR (MC192; 1:10; Alomone Labs, Jerusalem, Israel).

    Techniques: Immunoprecipitation, Transfection, Expressing

    Necdin enhances NGF/TrkA signaling in PC12 cells. A , Coimmunoprecipitation assay for the association between TrkA and p75NTR. PC12 cells infected with recombinant adenoviruses expressing necdin (Ad-Necdin) and β-galactosidase (Ad-LacZ) were treated with NGF (50 ng/ml) for 38 h. Cell lysates (400 μg) were immunoprecipitated (IP) with mouse anti-p75NTR antibody MC192 (αp75NTRm) and immunoblotted (IB) with anti-TrkA antibody (top panel). Expressed proteins in cell lysates are shown in the bottom panels. B , Neurite outgrowth assay. Ad-Necdin- and Ad-LacZ-infected PC12 cells were treated with NGF for 38 h, and cells bearing extended neurites were counted (mean ± SEM; n = 4; p

    Journal: The Journal of Neuroscience

    Article Title: Disruption of the Paternal Necdin Gene Diminishes TrkA Signaling for Sensory Neuron Survival

    doi: 10.1523/JNEUROSCI.2083-05.2005

    Figure Lengend Snippet: Necdin enhances NGF/TrkA signaling in PC12 cells. A , Coimmunoprecipitation assay for the association between TrkA and p75NTR. PC12 cells infected with recombinant adenoviruses expressing necdin (Ad-Necdin) and β-galactosidase (Ad-LacZ) were treated with NGF (50 ng/ml) for 38 h. Cell lysates (400 μg) were immunoprecipitated (IP) with mouse anti-p75NTR antibody MC192 (αp75NTRm) and immunoblotted (IB) with anti-TrkA antibody (top panel). Expressed proteins in cell lysates are shown in the bottom panels. B , Neurite outgrowth assay. Ad-Necdin- and Ad-LacZ-infected PC12 cells were treated with NGF for 38 h, and cells bearing extended neurites were counted (mean ± SEM; n = 4; p

    Article Snippet: Antibodies used for coimmunoprecipitation assay are as follows: FLAG (M2; 1:50; Sigma), Necdin (NC243; 1:50), and p75NTR (MC192; 1:10; Alomone Labs, Jerusalem, Israel).

    Techniques: Co-Immunoprecipitation Assay, Infection, Recombinant, Expressing, Immunoprecipitation, Neurite Outgrowth Assay

    Necdin expression is absent in DRGs of mutant mice lacking the paternal necdin gene. A , The restriction map of mouse Ndn locus. A 7 kb Ndn sequence containing coding (black) and 3′,5′ noncoding (hatched) regions was subcloned into the targeting vector. The 1.5 kb expression unit containing the pgk promoter with Neo r gene (Pgk1/neo r ), polyadenylation signal [Poly(A) signal], was inserted to disrupt the Ndn coding sequence. The vector was introduced to TT2 cells for homologous recombination as described in Materials and Methods. B , Absence of necdin expression in the brain and spinal cord of mice deficient in the paternal necdin gene. Frozen sections of the hypothalamus and spinal cord from E13.5 wild-type (Ndn +m/+p ) and necdin-deficient (Ndn +m/-p ) littermates were immunostained for necdin. HT, Hypothalamus; SC, spinal cord; DBB, diagonal band of Broca; ML, mantle layer. Scale bars: HT, 50 μm; SC, 100 μm. C , Expression patterns of TrkA and p75NTR in necdin-deficient DRGs. Frozen sections of cervical DRGs from E12.5 wild-type (Ndn +m/+p ) and necdin-deficient (Ndn +m/-p ) littermates were immunostained for necdin, TrkA, and p75NTR. Scale bar, 50 μm. D , E , Western blot analysis of necdin, TrkA, p75NTR, and phosphorylated MAPK. Equal amounts of DRG lysates from E12.5 embryos were analyzed by Western blotting. Each lane represents pooled DRGs from a single embryo. Molecular sizes in kilodaltons are shown to the right. E , Signal intensities of TrkA, p75NTR, and pMAPK shown in D were normalized to those of tubulin (TB) and MAPK (mean ± SEM; n = 5 for each protein). * p

    Journal: The Journal of Neuroscience

    Article Title: Disruption of the Paternal Necdin Gene Diminishes TrkA Signaling for Sensory Neuron Survival

    doi: 10.1523/JNEUROSCI.2083-05.2005

    Figure Lengend Snippet: Necdin expression is absent in DRGs of mutant mice lacking the paternal necdin gene. A , The restriction map of mouse Ndn locus. A 7 kb Ndn sequence containing coding (black) and 3′,5′ noncoding (hatched) regions was subcloned into the targeting vector. The 1.5 kb expression unit containing the pgk promoter with Neo r gene (Pgk1/neo r ), polyadenylation signal [Poly(A) signal], was inserted to disrupt the Ndn coding sequence. The vector was introduced to TT2 cells for homologous recombination as described in Materials and Methods. B , Absence of necdin expression in the brain and spinal cord of mice deficient in the paternal necdin gene. Frozen sections of the hypothalamus and spinal cord from E13.5 wild-type (Ndn +m/+p ) and necdin-deficient (Ndn +m/-p ) littermates were immunostained for necdin. HT, Hypothalamus; SC, spinal cord; DBB, diagonal band of Broca; ML, mantle layer. Scale bars: HT, 50 μm; SC, 100 μm. C , Expression patterns of TrkA and p75NTR in necdin-deficient DRGs. Frozen sections of cervical DRGs from E12.5 wild-type (Ndn +m/+p ) and necdin-deficient (Ndn +m/-p ) littermates were immunostained for necdin, TrkA, and p75NTR. Scale bar, 50 μm. D , E , Western blot analysis of necdin, TrkA, p75NTR, and phosphorylated MAPK. Equal amounts of DRG lysates from E12.5 embryos were analyzed by Western blotting. Each lane represents pooled DRGs from a single embryo. Molecular sizes in kilodaltons are shown to the right. E , Signal intensities of TrkA, p75NTR, and pMAPK shown in D were normalized to those of tubulin (TB) and MAPK (mean ± SEM; n = 5 for each protein). * p

    Article Snippet: Antibodies used for coimmunoprecipitation assay are as follows: FLAG (M2; 1:50; Sigma), Necdin (NC243; 1:50), and p75NTR (MC192; 1:10; Alomone Labs, Jerusalem, Israel).

    Techniques: Expressing, Mutagenesis, Mouse Assay, Sequencing, Plasmid Preparation, Homologous Recombination, Western Blot

    Effect of 8-OH-DPAT on the protein expression of molecules from the BDNF signaling pathway in the HC, determined by Western Blot. (A) mBDNF. (B) TrkB. (C) proBDNF. (D) p75 in hyposerotonergic (PCPA-treated) or control (vehicle-treated) mice receiving the 5-HT 1 A agonist 8-OH-DPAT (striped bars) or NaCl (white bars). (E) Representative membrane showing signal for p75 (75 kDa) and tubuline (52 kDa). 1: Control-NaCl. 2: Control-DPAT. 3: PCPA-NaCl. 4: PCPA-DPAT. Data are expressed as mean ± S.E.M., n = 6 per experimental group. ∗ p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Differential Hippocampal Expression of BDNF Isoforms and Their Receptors Under Diverse Configurations of the Serotonergic System in a Mice Model of Increased Neuronal Survival

    doi: 10.3389/fncel.2019.00384

    Figure Lengend Snippet: Effect of 8-OH-DPAT on the protein expression of molecules from the BDNF signaling pathway in the HC, determined by Western Blot. (A) mBDNF. (B) TrkB. (C) proBDNF. (D) p75 in hyposerotonergic (PCPA-treated) or control (vehicle-treated) mice receiving the 5-HT 1 A agonist 8-OH-DPAT (striped bars) or NaCl (white bars). (E) Representative membrane showing signal for p75 (75 kDa) and tubuline (52 kDa). 1: Control-NaCl. 2: Control-DPAT. 3: PCPA-NaCl. 4: PCPA-DPAT. Data are expressed as mean ± S.E.M., n = 6 per experimental group. ∗ p

    Article Snippet: Western Blotting Membranes were incubated for 1 h with blocking solution (5% milk in TBST) and then probed overnight at 4°C with mouse anti-BDNF (1:2000; Icosagen; 327-100 clone 3C11), rabbit anti-p75 (1:700; Alomone Labs; ANT-007), rabbit anti-TrkB (1:700; Alomone Labs; ANT-019), and rabbit anti-proBDNF (1:250; Abcam; ab72440) in TBST. β-III Tubulin was used as a loading control (1:2500; R & D Systems).

    Techniques: Expressing, Western Blot, Mouse Assay

    Protein expression of molecules from the BDNF signaling pathway determined by Western Blot and representative membranes. (A) mBDNF. (B) TrkB. (C) proBDNF. (D) p75 in hyposerotonergic (PCPA-treated) or control (vehicle-treated) mice for 4 weeks. (E–H) Representative membrane showing signal for TrkB, p75, pro-BDNF, and mBDNF in Control and PCPA-treatd mice. Data are expressed as mean ± S.E.M., n = 11 (control) and 12 (PCPA). ∗ p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Differential Hippocampal Expression of BDNF Isoforms and Their Receptors Under Diverse Configurations of the Serotonergic System in a Mice Model of Increased Neuronal Survival

    doi: 10.3389/fncel.2019.00384

    Figure Lengend Snippet: Protein expression of molecules from the BDNF signaling pathway determined by Western Blot and representative membranes. (A) mBDNF. (B) TrkB. (C) proBDNF. (D) p75 in hyposerotonergic (PCPA-treated) or control (vehicle-treated) mice for 4 weeks. (E–H) Representative membrane showing signal for TrkB, p75, pro-BDNF, and mBDNF in Control and PCPA-treatd mice. Data are expressed as mean ± S.E.M., n = 11 (control) and 12 (PCPA). ∗ p

    Article Snippet: Western Blotting Membranes were incubated for 1 h with blocking solution (5% milk in TBST) and then probed overnight at 4°C with mouse anti-BDNF (1:2000; Icosagen; 327-100 clone 3C11), rabbit anti-p75 (1:700; Alomone Labs; ANT-007), rabbit anti-TrkB (1:700; Alomone Labs; ANT-019), and rabbit anti-proBDNF (1:250; Abcam; ab72440) in TBST. β-III Tubulin was used as a loading control (1:2500; R & D Systems).

    Techniques: Expressing, Western Blot, Mouse Assay

    Suggested model. In order to arrive at the cell body and subsequently the CNS, rabies virus hijacks a fast route using the p75NTR endosomal pathway. In a p75NTR dependent path, RABV manipulates the axonal transport machinery to migrate faster to the cell body. An alternative, slower path, may involve alternative RABV receptors.

    Journal: PLoS Pathogens

    Article Title: Rabies Virus Hijacks and Accelerates the p75NTR Retrograde Axonal Transport Machinery

    doi: 10.1371/journal.ppat.1004348

    Figure Lengend Snippet: Suggested model. In order to arrive at the cell body and subsequently the CNS, rabies virus hijacks a fast route using the p75NTR endosomal pathway. In a p75NTR dependent path, RABV manipulates the axonal transport machinery to migrate faster to the cell body. An alternative, slower path, may involve alternative RABV receptors.

    Article Snippet: For RABV-p75 imaging, explant cultures were incubated with fluorescent anti-p75NTR (ANT-007-AO, Alomone Labs) for 10 minutes and washed 3 times in poor neurobasal medium prior to imaging.

    Techniques:

    RABV is retrogradely transported with neurotrophin receptors. ( A–D ), Retrograde transport of EGFP-RABV, added to the distal axon compartment of DRG explant previously treated with fluorescent antibodies against p75NTR and TrkA. Arrowheads: RABV puncta positive for p75NTR, arrows: RABV puncta positive for p75NTR and TrkA. Scale bar = 10 µm. ( E,F ) Co-localisation of RABV with p75NTR and TrkA calculated from two and one experiments, respectively. ( G ) Trajectories of RABV trafficked with neurotrophin receptors (NTFR, Blue) or without (Red), illustrating a more processive displacement over time of RABV with NTFR. ( H ) Merged kymographs of RABV (green) p75NTR (red) and TrkA (cyan), drawn for multi-channel time lapse. Vertical scale bar = 5 µm, horizontal scale bar = 40 seconds.

    Journal: PLoS Pathogens

    Article Title: Rabies Virus Hijacks and Accelerates the p75NTR Retrograde Axonal Transport Machinery

    doi: 10.1371/journal.ppat.1004348

    Figure Lengend Snippet: RABV is retrogradely transported with neurotrophin receptors. ( A–D ), Retrograde transport of EGFP-RABV, added to the distal axon compartment of DRG explant previously treated with fluorescent antibodies against p75NTR and TrkA. Arrowheads: RABV puncta positive for p75NTR, arrows: RABV puncta positive for p75NTR and TrkA. Scale bar = 10 µm. ( E,F ) Co-localisation of RABV with p75NTR and TrkA calculated from two and one experiments, respectively. ( G ) Trajectories of RABV trafficked with neurotrophin receptors (NTFR, Blue) or without (Red), illustrating a more processive displacement over time of RABV with NTFR. ( H ) Merged kymographs of RABV (green) p75NTR (red) and TrkA (cyan), drawn for multi-channel time lapse. Vertical scale bar = 5 µm, horizontal scale bar = 40 seconds.

    Article Snippet: For RABV-p75 imaging, explant cultures were incubated with fluorescent anti-p75NTR (ANT-007-AO, Alomone Labs) for 10 minutes and washed 3 times in poor neurobasal medium prior to imaging.

    Techniques:

    RABV binds and internalizes with p75NTR in DRG neuron tips. Co-localization of EGFP-RABV with p75NTR is shown by live TIRF imaging and sub-pixel localization algorithms. ( A ) RABV-p75 particles shift from the periphery to the center of the growth cone, where they are internalized into the cell. Lower panels zoom in on dashed square, showing co-localized puncta (left) shifting towards the center of the growth cone (middle) until finally internalized (right). ( B ) Presentation of six separate events of RABV and p75NTR binding and internalization on the surface of the growth cone shown in (A). Colored trajectories denote displacement from point of detection to point of disappearance. ( C ) RABV and p75NTR are internalized together, illustrated by corresponding plots of puncta intensity over time (normalized to background), calculated for co-localized particles shown in lower panels of (A). Scale bars = 5 µm. ( D ) Zoom-in on colocalized RABV and p75 spot, taken from panel (A), scale bar = 1 µm. ( E ) Overlay of 1D-Gaussian fits of p75 and RABV intensity profiles at the x-axis of the image in panel (D). ( F ) Representative overlay of radial symmetry fits of the x-y intensity profiles of p75 and RABV spots. σ is the standard deviation of each fitting function; distance between the two spot centers is 51.3 nm. ( G ) Knockdown of p75NTR decreases rabies virus infection for shorts time incubation. DRGs embryonic cells infected with lentiviral vectors (LV) containing 4 different EGFP-tagged shRNA's against p75NTR or LV-EGFP, were transfected with RABV for 30 or 120 minutes. Low levels of infected neurons were found in shRNA-p75-EGFP cells Average RABV infection rates were normalized to LV-EGFP controls (n = 4 experiments, error bars = SEM, *p

    Journal: PLoS Pathogens

    Article Title: Rabies Virus Hijacks and Accelerates the p75NTR Retrograde Axonal Transport Machinery

    doi: 10.1371/journal.ppat.1004348

    Figure Lengend Snippet: RABV binds and internalizes with p75NTR in DRG neuron tips. Co-localization of EGFP-RABV with p75NTR is shown by live TIRF imaging and sub-pixel localization algorithms. ( A ) RABV-p75 particles shift from the periphery to the center of the growth cone, where they are internalized into the cell. Lower panels zoom in on dashed square, showing co-localized puncta (left) shifting towards the center of the growth cone (middle) until finally internalized (right). ( B ) Presentation of six separate events of RABV and p75NTR binding and internalization on the surface of the growth cone shown in (A). Colored trajectories denote displacement from point of detection to point of disappearance. ( C ) RABV and p75NTR are internalized together, illustrated by corresponding plots of puncta intensity over time (normalized to background), calculated for co-localized particles shown in lower panels of (A). Scale bars = 5 µm. ( D ) Zoom-in on colocalized RABV and p75 spot, taken from panel (A), scale bar = 1 µm. ( E ) Overlay of 1D-Gaussian fits of p75 and RABV intensity profiles at the x-axis of the image in panel (D). ( F ) Representative overlay of radial symmetry fits of the x-y intensity profiles of p75 and RABV spots. σ is the standard deviation of each fitting function; distance between the two spot centers is 51.3 nm. ( G ) Knockdown of p75NTR decreases rabies virus infection for shorts time incubation. DRGs embryonic cells infected with lentiviral vectors (LV) containing 4 different EGFP-tagged shRNA's against p75NTR or LV-EGFP, were transfected with RABV for 30 or 120 minutes. Low levels of infected neurons were found in shRNA-p75-EGFP cells Average RABV infection rates were normalized to LV-EGFP controls (n = 4 experiments, error bars = SEM, *p

    Article Snippet: For RABV-p75 imaging, explant cultures were incubated with fluorescent anti-p75NTR (ANT-007-AO, Alomone Labs) for 10 minutes and washed 3 times in poor neurobasal medium prior to imaging.

    Techniques: Imaging, Binding Assay, Standard Deviation, Infection, Incubation, shRNA, Transfection

    RABV travels faster and is more directed when transported with p75NTR. ( A–C ) Multi-channel live imaging of EGFP-RABV 2 hours after addition to distal axon compartment of DRG explant previously treated with a fluorescent antibody against p75NTR. Arrowheads: p75NTR-positive RABV puncta, scale bar = 10 µm. ( D,E ) Kymographs of and P75NTR extracted from time lapse depicted in (A–C). ( F ) RABV-only tracks (green) are less directed than RABV-p75NTR tracks (yellow), as shown when overlaying corresponding kymographs. Vertical scale bar = 5 µm, horizontal scale bar = 40 seconds. ( G–O ) Characterization of directed RABV puncta, transported with and without p75NTR, n = 184 and n = 122, respectively. (G) RABV presents higher speeds when transported with p75NTR, due to less frequent (H) and shorter pauses (I). Overall RABV-p75NTR spent less time paused on average (J), Diameter and intensity measurements revealed that RABV puncta positive for p75NTR were larger ( K ) and had higher intensity levels ( L ) than p75NTR-negative puncta. ( M–O ) p75NTR positive puncta (blue) are faster, more directed and present higher displacements over time, compared to p75NTR negative puncta (red), illustrated by distribution of instantaneous velocities in (M) (RABV+p75: n = 8051 events; RABV-p75: n = 7423 events) displacement plotted over time (N) and mean square displacement (O). Data is pulled from two separate experiments, error bars represent SEM. *p

    Journal: PLoS Pathogens

    Article Title: Rabies Virus Hijacks and Accelerates the p75NTR Retrograde Axonal Transport Machinery

    doi: 10.1371/journal.ppat.1004348

    Figure Lengend Snippet: RABV travels faster and is more directed when transported with p75NTR. ( A–C ) Multi-channel live imaging of EGFP-RABV 2 hours after addition to distal axon compartment of DRG explant previously treated with a fluorescent antibody against p75NTR. Arrowheads: p75NTR-positive RABV puncta, scale bar = 10 µm. ( D,E ) Kymographs of and P75NTR extracted from time lapse depicted in (A–C). ( F ) RABV-only tracks (green) are less directed than RABV-p75NTR tracks (yellow), as shown when overlaying corresponding kymographs. Vertical scale bar = 5 µm, horizontal scale bar = 40 seconds. ( G–O ) Characterization of directed RABV puncta, transported with and without p75NTR, n = 184 and n = 122, respectively. (G) RABV presents higher speeds when transported with p75NTR, due to less frequent (H) and shorter pauses (I). Overall RABV-p75NTR spent less time paused on average (J), Diameter and intensity measurements revealed that RABV puncta positive for p75NTR were larger ( K ) and had higher intensity levels ( L ) than p75NTR-negative puncta. ( M–O ) p75NTR positive puncta (blue) are faster, more directed and present higher displacements over time, compared to p75NTR negative puncta (red), illustrated by distribution of instantaneous velocities in (M) (RABV+p75: n = 8051 events; RABV-p75: n = 7423 events) displacement plotted over time (N) and mean square displacement (O). Data is pulled from two separate experiments, error bars represent SEM. *p

    Article Snippet: For RABV-p75 imaging, explant cultures were incubated with fluorescent anti-p75NTR (ANT-007-AO, Alomone Labs) for 10 minutes and washed 3 times in poor neurobasal medium prior to imaging.

    Techniques: Imaging