cleavage resistant prongf  (Alomone Labs)


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    Alomone Labs cleavage resistant prongf
    <t>Overexpression</t> of <t>proNGF</t> induced glial Müller activation and inflammation. Immunohistochemical analysis of retinas was performed 4 weeks after intravitreal injections of pGFP alone (control) ( a–e ), proNGF plus scrambled p75 NTR shRNA (proNGF+Scr)
    Cleavage Resistant Prongf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cleavage resistant prongf/product/Alomone Labs
    Average 94 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    cleavage resistant prongf - by Bioz Stars, 2022-09
    94/100 stars

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    1) Product Images from "Modulation of p75NTR prevents diabetes- and proNGF-induced retinal inflammation and blood–retina barrier breakdown in mice and rats"

    Article Title: Modulation of p75NTR prevents diabetes- and proNGF-induced retinal inflammation and blood–retina barrier breakdown in mice and rats

    Journal: Diabetologia

    doi: 10.1007/s00125-013-2998-6

    Overexpression of proNGF induced glial Müller activation and inflammation. Immunohistochemical analysis of retinas was performed 4 weeks after intravitreal injections of pGFP alone (control) ( a–e ), proNGF plus scrambled p75 NTR shRNA (proNGF+Scr)
    Figure Legend Snippet: Overexpression of proNGF induced glial Müller activation and inflammation. Immunohistochemical analysis of retinas was performed 4 weeks after intravitreal injections of pGFP alone (control) ( a–e ), proNGF plus scrambled p75 NTR shRNA (proNGF+Scr)

    Techniques Used: Over Expression, Activation Assay, Immunohistochemistry, shRNA

    2) Product Images from "Modulation of p75NTR prevents diabetes- and proNGF-induced retinal inflammation and blood–retina barrier breakdown in mice and rats"

    Article Title: Modulation of p75NTR prevents diabetes- and proNGF-induced retinal inflammation and blood–retina barrier breakdown in mice and rats

    Journal: Diabetologia

    doi: 10.1007/s00125-013-2998-6

    Overexpression of proNGF induced glial Müller activation and inflammation. Immunohistochemical analysis of retinas was performed 4 weeks after intravitreal injections of pGFP alone (control) ( a–e ), proNGF plus scrambled p75 NTR shRNA (proNGF+Scr)
    Figure Legend Snippet: Overexpression of proNGF induced glial Müller activation and inflammation. Immunohistochemical analysis of retinas was performed 4 weeks after intravitreal injections of pGFP alone (control) ( a–e ), proNGF plus scrambled p75 NTR shRNA (proNGF+Scr)

    Techniques Used: Over Expression, Activation Assay, Immunohistochemistry, shRNA

    3) Product Images from "p75NTR is highly expressed in vestibular schwannomas and promotes cell survival by activating NF-κB"

    Article Title: p75NTR is highly expressed in vestibular schwannomas and promotes cell survival by activating NF-κB

    Journal: Glia

    doi: 10.1002/glia.22709

    ProNGF activates JNK and NF-κB in primary VS cultures. A . Western blots of protein lysates from primary VS cultures treated Ad5.empty vector (Ad.EV) or Ad5.merlin (Ad.merlin) and probed with anti-phosphorylated JNK antibodies. The blots were stripped
    Figure Legend Snippet: ProNGF activates JNK and NF-κB in primary VS cultures. A . Western blots of protein lysates from primary VS cultures treated Ad5.empty vector (Ad.EV) or Ad5.merlin (Ad.merlin) and probed with anti-phosphorylated JNK antibodies. The blots were stripped

    Techniques Used: Western Blot, Plasmid Preparation

    ProNGF protects VS cells from apoptosis due to JNK inhibition. A-D . Primary VS cultures were maintained in the presence of the JNK inhibitors, I-JIP or SP600125, with or without treatment with proNGF (3 nM) (A) of proBDNF (3 nM) (D). Cultures were immunostained
    Figure Legend Snippet: ProNGF protects VS cells from apoptosis due to JNK inhibition. A-D . Primary VS cultures were maintained in the presence of the JNK inhibitors, I-JIP or SP600125, with or without treatment with proNGF (3 nM) (A) of proBDNF (3 nM) (D). Cultures were immunostained

    Techniques Used: Inhibition

    4) Product Images from "p75NTR is highly expressed in vestibular schwannomas and promotes cell survival by activating NF-κB"

    Article Title: p75NTR is highly expressed in vestibular schwannomas and promotes cell survival by activating NF-κB

    Journal: Glia

    doi: 10.1002/glia.22709

    ProNGF fails to induce apoptosis in primary vestibular schwannoma cultures. A . Primary VS cultures immunostained with anti-p75 NTR antibody (green). Nuclei are labeled with DAPI (blue). B,C . Primary human VS cultures were treated with escalating doses
    Figure Legend Snippet: ProNGF fails to induce apoptosis in primary vestibular schwannoma cultures. A . Primary VS cultures immunostained with anti-p75 NTR antibody (green). Nuclei are labeled with DAPI (blue). B,C . Primary human VS cultures were treated with escalating doses

    Techniques Used: Labeling

    ProNGF protects VS cells from apoptosis due to JNK inhibition. A-D . Primary VS cultures were maintained in the presence of the JNK inhibitors, I-JIP or SP600125, with or without treatment with proNGF (3 nM) (A) of proBDNF (3 nM) (D). Cultures were immunostained
    Figure Legend Snippet: ProNGF protects VS cells from apoptosis due to JNK inhibition. A-D . Primary VS cultures were maintained in the presence of the JNK inhibitors, I-JIP or SP600125, with or without treatment with proNGF (3 nM) (A) of proBDNF (3 nM) (D). Cultures were immunostained

    Techniques Used: Inhibition

    5) Product Images from "Modulation of p75NTR prevents diabetes- and proNGF-induced retinal inflammation and blood–retina barrier breakdown in mice and rats"

    Article Title: Modulation of p75NTR prevents diabetes- and proNGF-induced retinal inflammation and blood–retina barrier breakdown in mice and rats

    Journal: Diabetologia

    doi: 10.1007/s00125-013-2998-6

    Modulation of p75 NTR action or expression prevents proNGF-mediated inflammation in Müller cells. ( a ) Representative blots and densitometric analysis of TNF-α ( b ), p-NFκB (black columns) and NFκB (white columns) ( c ), and
    Figure Legend Snippet: Modulation of p75 NTR action or expression prevents proNGF-mediated inflammation in Müller cells. ( a ) Representative blots and densitometric analysis of TNF-α ( b ), p-NFκB (black columns) and NFκB (white columns) ( c ), and

    Techniques Used: Expressing

    Overexpression of proNGF induced glial Müller activation and inflammation. Immunohistochemical analysis of retinas was performed 4 weeks after intravitreal injections of pGFP alone (control) ( a–e ), proNGF plus scrambled p75 NTR shRNA (proNGF+Scr)
    Figure Legend Snippet: Overexpression of proNGF induced glial Müller activation and inflammation. Immunohistochemical analysis of retinas was performed 4 weeks after intravitreal injections of pGFP alone (control) ( a–e ), proNGF plus scrambled p75 NTR shRNA (proNGF+Scr)

    Techniques Used: Over Expression, Activation Assay, Immunohistochemistry, shRNA

    6) Product Images from "ProNGF-p75NTR axis plays a proinflammatory role in inflamed joints: a novel pathogenic mechanism in chronic arthritis"

    Article Title: ProNGF-p75NTR axis plays a proinflammatory role in inflamed joints: a novel pathogenic mechanism in chronic arthritis

    Journal: RMD Open

    doi: 10.1136/rmdopen-2017-000441

    Mature and immature NGF forms in patients with JIA and RA. (A,B) Only proNGF forms of different molecular weights but not mature NGF were detected by western blot in synovial fluids of 12 patients with JIA (40 μg total protein). (B) Blocking the anti-NGF antibody by adding mature NGF results in the disappearance of the specific proNGF bands observed in synovial fluids of three patients with JIA, as well as of the band of the 25 kDa commercial proNGF that was added as positive control. (C) In JIA and RA synovial fluids, proNGF and mature NGF (mNGF) concentrations were assessed using a newly developed ELISA showing that proNGF is 4.8-fold higher in JIA (n=27) and 16.8-fold higher in patients with RA (n=5) than mNGF. (D) Real-time PCR shows a very low expression of NGF mRNA in mononuclear cells obtained from peripheral blood of healthy donors (CTRL PBMC) or in mononuclear cells from peripheral blood (JIA PBMC) and from synovial fluids of patients with JIA (JIA SFMC). High NGF mRNA expression levels characterised synovial tissues (RA synovia; n=5). Fibroblast-like synoviocytes (FLS; n=3) of patients with RA express more NGF mRNA than FLS from osteoarthritis patients (OA FLS) (n=4) and control fibroblasts (CTRL FB; n=4). Results were expressed as arbitrary units (AU) and obtained after normalisation with the housekeeping gene glyceraldehyde 3-phosphate dehydrogenase (GAPDH). (E) Both unstimulated (US) and LPS-stimulated JIA SFMC do not release proNGF or mNGF in the supernatants. Commercial proNGF and mNGF were added as positive controls. (F) Western blot shows the stability of exogenous added mNGF and proNGF. After its addition to culture media, proNGF is still detected after 3 hours and undergoes a minor maturation into mNGF in the first 2 hours of incubation (left side of F). mNGF is not degraded in the first 3 hours after its addition (right side of F). Commercial proNGF and mNGF were included as positive controls. (G) Neither proNGF nor mNGF were released in culture media of US or LPS-stimulated JIA SFMC after 18 hours of incubation. Exogenous proNGF or mNGF were detected only at time of supplementation (in agreement with F) but were no longer detectable after 18 hours of incubation. Commercial proNGF and mNGF were included as positive controls. (H) ELISA instead shows that conditioned media of RA FLS (n=5) have higher concentrations of proNGF than control fibroblasts (CTRL FB; n=5) and that the concentration of proNGF is higher than mNGF after 18 hours of incubation. *p
    Figure Legend Snippet: Mature and immature NGF forms in patients with JIA and RA. (A,B) Only proNGF forms of different molecular weights but not mature NGF were detected by western blot in synovial fluids of 12 patients with JIA (40 μg total protein). (B) Blocking the anti-NGF antibody by adding mature NGF results in the disappearance of the specific proNGF bands observed in synovial fluids of three patients with JIA, as well as of the band of the 25 kDa commercial proNGF that was added as positive control. (C) In JIA and RA synovial fluids, proNGF and mature NGF (mNGF) concentrations were assessed using a newly developed ELISA showing that proNGF is 4.8-fold higher in JIA (n=27) and 16.8-fold higher in patients with RA (n=5) than mNGF. (D) Real-time PCR shows a very low expression of NGF mRNA in mononuclear cells obtained from peripheral blood of healthy donors (CTRL PBMC) or in mononuclear cells from peripheral blood (JIA PBMC) and from synovial fluids of patients with JIA (JIA SFMC). High NGF mRNA expression levels characterised synovial tissues (RA synovia; n=5). Fibroblast-like synoviocytes (FLS; n=3) of patients with RA express more NGF mRNA than FLS from osteoarthritis patients (OA FLS) (n=4) and control fibroblasts (CTRL FB; n=4). Results were expressed as arbitrary units (AU) and obtained after normalisation with the housekeeping gene glyceraldehyde 3-phosphate dehydrogenase (GAPDH). (E) Both unstimulated (US) and LPS-stimulated JIA SFMC do not release proNGF or mNGF in the supernatants. Commercial proNGF and mNGF were added as positive controls. (F) Western blot shows the stability of exogenous added mNGF and proNGF. After its addition to culture media, proNGF is still detected after 3 hours and undergoes a minor maturation into mNGF in the first 2 hours of incubation (left side of F). mNGF is not degraded in the first 3 hours after its addition (right side of F). Commercial proNGF and mNGF were included as positive controls. (G) Neither proNGF nor mNGF were released in culture media of US or LPS-stimulated JIA SFMC after 18 hours of incubation. Exogenous proNGF or mNGF were detected only at time of supplementation (in agreement with F) but were no longer detectable after 18 hours of incubation. Commercial proNGF and mNGF were included as positive controls. (H) ELISA instead shows that conditioned media of RA FLS (n=5) have higher concentrations of proNGF than control fibroblasts (CTRL FB; n=5) and that the concentration of proNGF is higher than mNGF after 18 hours of incubation. *p

    Techniques Used: Western Blot, Blocking Assay, Positive Control, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Expressing, Incubation, Concentration Assay

    proNGF and NGF activated different intracellular pathways. (A) The neutralisation of p75NTR using either a specific anti-p75NTR antibody (a-p75; 2.5 µg/mL) or using the specific pharmacological inhibitor LM11A.31 (10 nM), decreased interleukin (IL)-6 release in LPS-activated SFMC of patients with JIA (n=16) treated with proNGF. On the contrary, the blocking of TrkA with a neutralising antibody (a-TrkA; 3 µg/mL) did not affect IL-6 release. (B) Western blot shows an increased phosphorylation of p38 and JNK after 5 minutes of proNGF stimulation in LPS-treated SFMC. mNGF treatment did not affect the phosphorylation of these downstream molecules. The result is representative of one out of three independent experiments. Results of the densitometric analysis of all experiments are expressed as arbitrary units (AU). (C) In LPS-activated SFMC of three different patients with JIA, preincubation with the inhibitor LM11A-31 (10 nM), which blocks the binding of proNGF to p75NTR, abolished the phosphorylation of p38 and JNK induced after 5 minutes of proNGF addition. Results of the densitometric analysis of all experiments are expressed as arbitrary units (AU). *p
    Figure Legend Snippet: proNGF and NGF activated different intracellular pathways. (A) The neutralisation of p75NTR using either a specific anti-p75NTR antibody (a-p75; 2.5 µg/mL) or using the specific pharmacological inhibitor LM11A.31 (10 nM), decreased interleukin (IL)-6 release in LPS-activated SFMC of patients with JIA (n=16) treated with proNGF. On the contrary, the blocking of TrkA with a neutralising antibody (a-TrkA; 3 µg/mL) did not affect IL-6 release. (B) Western blot shows an increased phosphorylation of p38 and JNK after 5 minutes of proNGF stimulation in LPS-treated SFMC. mNGF treatment did not affect the phosphorylation of these downstream molecules. The result is representative of one out of three independent experiments. Results of the densitometric analysis of all experiments are expressed as arbitrary units (AU). (C) In LPS-activated SFMC of three different patients with JIA, preincubation with the inhibitor LM11A-31 (10 nM), which blocks the binding of proNGF to p75NTR, abolished the phosphorylation of p38 and JNK induced after 5 minutes of proNGF addition. Results of the densitometric analysis of all experiments are expressed as arbitrary units (AU). *p

    Techniques Used: Blocking Assay, Western Blot, Binding Assay

    Ex vivo effects of proNGF and NGF on JIA mononuclear cells. (A) No changes in cell viability were observed when mononuclear cells from synovial fluid (SFMC) of patients with JIA were treated for 24 hours with proNGF or mNGF, with or without LPS stimulation. Apoptosis was assessed by Annexin V flow cytometry analysis. The results represent one of three independent experiments performed in duplicate. (B) While proNGF addition alone did not induce the production of interleukin (IL)-6 in unstimulated SFMC, in LPS-activated cells proNGF induced IL-6 release in a dose-dependent manner with a maximal effect at 200 ng/mL (n=4). (C) proNGF stimulation significantly increased IL-6, IL-1β, IL-8 and tumor necrosis factor-α (TNF-α) mRNA expression levels in LPS-stimulated SFMC (n=18) of patients with JIA after 3 hours of incubation, but did not modify IL-10 mRNA expression in either CTRL or JIA mononuclear cells. Mature NGF treatment did not significantly modify proinflammatory cytokine levels or IL-10 mRNA expression in JIA mononuclear cells, while it increased IL-10 mRNA levels in CTRL PBMC. Results are expressed as arbitrary units (AU) and obtained after normalisation with the housekeeping gene GAPDH. (D) proNGF treatment significantly increased IL-6 release after 18 hours (measured by ELISA) in LPS-stimulated SFMC from patients with JIA (n=16) compared with PBMC of healthy CTRL (n=6). Mature NGF treatment did not affect IL-6 protein levels. *p
    Figure Legend Snippet: Ex vivo effects of proNGF and NGF on JIA mononuclear cells. (A) No changes in cell viability were observed when mononuclear cells from synovial fluid (SFMC) of patients with JIA were treated for 24 hours with proNGF or mNGF, with or without LPS stimulation. Apoptosis was assessed by Annexin V flow cytometry analysis. The results represent one of three independent experiments performed in duplicate. (B) While proNGF addition alone did not induce the production of interleukin (IL)-6 in unstimulated SFMC, in LPS-activated cells proNGF induced IL-6 release in a dose-dependent manner with a maximal effect at 200 ng/mL (n=4). (C) proNGF stimulation significantly increased IL-6, IL-1β, IL-8 and tumor necrosis factor-α (TNF-α) mRNA expression levels in LPS-stimulated SFMC (n=18) of patients with JIA after 3 hours of incubation, but did not modify IL-10 mRNA expression in either CTRL or JIA mononuclear cells. Mature NGF treatment did not significantly modify proinflammatory cytokine levels or IL-10 mRNA expression in JIA mononuclear cells, while it increased IL-10 mRNA levels in CTRL PBMC. Results are expressed as arbitrary units (AU) and obtained after normalisation with the housekeeping gene GAPDH. (D) proNGF treatment significantly increased IL-6 release after 18 hours (measured by ELISA) in LPS-stimulated SFMC from patients with JIA (n=16) compared with PBMC of healthy CTRL (n=6). Mature NGF treatment did not affect IL-6 protein levels. *p

    Techniques Used: Ex Vivo, Flow Cytometry, Cytometry, Expressing, Incubation, Enzyme-linked Immunosorbent Assay

    7) Product Images from "ProNGF-p75NTR axis plays a proinflammatory role in inflamed joints: a novel pathogenic mechanism in chronic arthritis"

    Article Title: ProNGF-p75NTR axis plays a proinflammatory role in inflamed joints: a novel pathogenic mechanism in chronic arthritis

    Journal: RMD Open

    doi: 10.1136/rmdopen-2017-000441

    Mature and immature NGF forms in patients with JIA and RA. (A,B) Only proNGF forms of different molecular weights but not mature NGF were detected by western blot in synovial fluids of 12 patients with JIA (40 μg total protein). (B) Blocking the anti-NGF antibody by adding mature NGF results in the disappearance of the specific proNGF bands observed in synovial fluids of three patients with JIA, as well as of the band of the 25 kDa commercial proNGF that was added as positive control. (C) In JIA and RA synovial fluids, proNGF and mature NGF (mNGF) concentrations were assessed using a newly developed ELISA showing that proNGF is 4.8-fold higher in JIA (n=27) and 16.8-fold higher in patients with RA (n=5) than mNGF. (D) Real-time PCR shows a very low expression of NGF mRNA in mononuclear cells obtained from peripheral blood of healthy donors (CTRL PBMC) or in mononuclear cells from peripheral blood (JIA PBMC) and from synovial fluids of patients with JIA (JIA SFMC). High NGF mRNA expression levels characterised synovial tissues (RA synovia; n=5). Fibroblast-like synoviocytes (FLS; n=3) of patients with RA express more NGF mRNA than FLS from osteoarthritis patients (OA FLS) (n=4) and control fibroblasts (CTRL FB; n=4). Results were expressed as arbitrary units (AU) and obtained after normalisation with the housekeeping gene glyceraldehyde 3-phosphate dehydrogenase (GAPDH). (E) Both unstimulated (US) and LPS-stimulated JIA SFMC do not release proNGF or mNGF in the supernatants. Commercial proNGF and mNGF were added as positive controls. (F) Western blot shows the stability of exogenous added mNGF and proNGF. After its addition to culture media, proNGF is still detected after 3 hours and undergoes a minor maturation into mNGF in the first 2 hours of incubation (left side of F). mNGF is not degraded in the first 3 hours after its addition (right side of F). Commercial proNGF and mNGF were included as positive controls. (G) Neither proNGF nor mNGF were released in culture media of US or LPS-stimulated JIA SFMC after 18 hours of incubation. Exogenous proNGF or mNGF were detected only at time of supplementation (in agreement with F) but were no longer detectable after 18 hours of incubation. Commercial proNGF and mNGF were included as positive controls. (H) ELISA instead shows that conditioned media of RA FLS (n=5) have higher concentrations of proNGF than control fibroblasts (CTRL FB; n=5) and that the concentration of proNGF is higher than mNGF after 18 hours of incubation. *p
    Figure Legend Snippet: Mature and immature NGF forms in patients with JIA and RA. (A,B) Only proNGF forms of different molecular weights but not mature NGF were detected by western blot in synovial fluids of 12 patients with JIA (40 μg total protein). (B) Blocking the anti-NGF antibody by adding mature NGF results in the disappearance of the specific proNGF bands observed in synovial fluids of three patients with JIA, as well as of the band of the 25 kDa commercial proNGF that was added as positive control. (C) In JIA and RA synovial fluids, proNGF and mature NGF (mNGF) concentrations were assessed using a newly developed ELISA showing that proNGF is 4.8-fold higher in JIA (n=27) and 16.8-fold higher in patients with RA (n=5) than mNGF. (D) Real-time PCR shows a very low expression of NGF mRNA in mononuclear cells obtained from peripheral blood of healthy donors (CTRL PBMC) or in mononuclear cells from peripheral blood (JIA PBMC) and from synovial fluids of patients with JIA (JIA SFMC). High NGF mRNA expression levels characterised synovial tissues (RA synovia; n=5). Fibroblast-like synoviocytes (FLS; n=3) of patients with RA express more NGF mRNA than FLS from osteoarthritis patients (OA FLS) (n=4) and control fibroblasts (CTRL FB; n=4). Results were expressed as arbitrary units (AU) and obtained after normalisation with the housekeeping gene glyceraldehyde 3-phosphate dehydrogenase (GAPDH). (E) Both unstimulated (US) and LPS-stimulated JIA SFMC do not release proNGF or mNGF in the supernatants. Commercial proNGF and mNGF were added as positive controls. (F) Western blot shows the stability of exogenous added mNGF and proNGF. After its addition to culture media, proNGF is still detected after 3 hours and undergoes a minor maturation into mNGF in the first 2 hours of incubation (left side of F). mNGF is not degraded in the first 3 hours after its addition (right side of F). Commercial proNGF and mNGF were included as positive controls. (G) Neither proNGF nor mNGF were released in culture media of US or LPS-stimulated JIA SFMC after 18 hours of incubation. Exogenous proNGF or mNGF were detected only at time of supplementation (in agreement with F) but were no longer detectable after 18 hours of incubation. Commercial proNGF and mNGF were included as positive controls. (H) ELISA instead shows that conditioned media of RA FLS (n=5) have higher concentrations of proNGF than control fibroblasts (CTRL FB; n=5) and that the concentration of proNGF is higher than mNGF after 18 hours of incubation. *p

    Techniques Used: Western Blot, Blocking Assay, Positive Control, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Expressing, Incubation, Concentration Assay

    proNGF and NGF activated different intracellular pathways. (A) The neutralisation of p75NTR using either a specific anti-p75NTR antibody (a-p75; 2.5 µg/mL) or using the specific pharmacological inhibitor LM11A.31 (10 nM), decreased interleukin (IL)-6 release in LPS-activated SFMC of patients with JIA (n=16) treated with proNGF. On the contrary, the blocking of TrkA with a neutralising antibody (a-TrkA; 3 µg/mL) did not affect IL-6 release. (B) Western blot shows an increased phosphorylation of p38 and JNK after 5 minutes of proNGF stimulation in LPS-treated SFMC. mNGF treatment did not affect the phosphorylation of these downstream molecules. The result is representative of one out of three independent experiments. Results of the densitometric analysis of all experiments are expressed as arbitrary units (AU). (C) In LPS-activated SFMC of three different patients with JIA, preincubation with the inhibitor LM11A-31 (10 nM), which blocks the binding of proNGF to p75NTR, abolished the phosphorylation of p38 and JNK induced after 5 minutes of proNGF addition. Results of the densitometric analysis of all experiments are expressed as arbitrary units (AU). *p
    Figure Legend Snippet: proNGF and NGF activated different intracellular pathways. (A) The neutralisation of p75NTR using either a specific anti-p75NTR antibody (a-p75; 2.5 µg/mL) or using the specific pharmacological inhibitor LM11A.31 (10 nM), decreased interleukin (IL)-6 release in LPS-activated SFMC of patients with JIA (n=16) treated with proNGF. On the contrary, the blocking of TrkA with a neutralising antibody (a-TrkA; 3 µg/mL) did not affect IL-6 release. (B) Western blot shows an increased phosphorylation of p38 and JNK after 5 minutes of proNGF stimulation in LPS-treated SFMC. mNGF treatment did not affect the phosphorylation of these downstream molecules. The result is representative of one out of three independent experiments. Results of the densitometric analysis of all experiments are expressed as arbitrary units (AU). (C) In LPS-activated SFMC of three different patients with JIA, preincubation with the inhibitor LM11A-31 (10 nM), which blocks the binding of proNGF to p75NTR, abolished the phosphorylation of p38 and JNK induced after 5 minutes of proNGF addition. Results of the densitometric analysis of all experiments are expressed as arbitrary units (AU). *p

    Techniques Used: Blocking Assay, Western Blot, Binding Assay

    Ex vivo effects of proNGF and NGF on JIA mononuclear cells. (A) No changes in cell viability were observed when mononuclear cells from synovial fluid (SFMC) of patients with JIA were treated for 24 hours with proNGF or mNGF, with or without LPS stimulation. Apoptosis was assessed by Annexin V flow cytometry analysis. The results represent one of three independent experiments performed in duplicate. (B) While proNGF addition alone did not induce the production of interleukin (IL)-6 in unstimulated SFMC, in LPS-activated cells proNGF induced IL-6 release in a dose-dependent manner with a maximal effect at 200 ng/mL (n=4). (C) proNGF stimulation significantly increased IL-6, IL-1β, IL-8 and tumor necrosis factor-α (TNF-α) mRNA expression levels in LPS-stimulated SFMC (n=18) of patients with JIA after 3 hours of incubation, but did not modify IL-10 mRNA expression in either CTRL or JIA mononuclear cells. Mature NGF treatment did not significantly modify proinflammatory cytokine levels or IL-10 mRNA expression in JIA mononuclear cells, while it increased IL-10 mRNA levels in CTRL PBMC. Results are expressed as arbitrary units (AU) and obtained after normalisation with the housekeeping gene GAPDH. (D) proNGF treatment significantly increased IL-6 release after 18 hours (measured by ELISA) in LPS-stimulated SFMC from patients with JIA (n=16) compared with PBMC of healthy CTRL (n=6). Mature NGF treatment did not affect IL-6 protein levels. *p
    Figure Legend Snippet: Ex vivo effects of proNGF and NGF on JIA mononuclear cells. (A) No changes in cell viability were observed when mononuclear cells from synovial fluid (SFMC) of patients with JIA were treated for 24 hours with proNGF or mNGF, with or without LPS stimulation. Apoptosis was assessed by Annexin V flow cytometry analysis. The results represent one of three independent experiments performed in duplicate. (B) While proNGF addition alone did not induce the production of interleukin (IL)-6 in unstimulated SFMC, in LPS-activated cells proNGF induced IL-6 release in a dose-dependent manner with a maximal effect at 200 ng/mL (n=4). (C) proNGF stimulation significantly increased IL-6, IL-1β, IL-8 and tumor necrosis factor-α (TNF-α) mRNA expression levels in LPS-stimulated SFMC (n=18) of patients with JIA after 3 hours of incubation, but did not modify IL-10 mRNA expression in either CTRL or JIA mononuclear cells. Mature NGF treatment did not significantly modify proinflammatory cytokine levels or IL-10 mRNA expression in JIA mononuclear cells, while it increased IL-10 mRNA levels in CTRL PBMC. Results are expressed as arbitrary units (AU) and obtained after normalisation with the housekeeping gene GAPDH. (D) proNGF treatment significantly increased IL-6 release after 18 hours (measured by ELISA) in LPS-stimulated SFMC from patients with JIA (n=16) compared with PBMC of healthy CTRL (n=6). Mature NGF treatment did not affect IL-6 protein levels. *p

    Techniques Used: Ex Vivo, Flow Cytometry, Cytometry, Expressing, Incubation, Enzyme-linked Immunosorbent Assay

    8) Product Images from "Modulation of p75NTR prevents diabetes- and proNGF-induced retinal inflammation and blood–retina barrier breakdown in mice and rats"

    Article Title: Modulation of p75NTR prevents diabetes- and proNGF-induced retinal inflammation and blood–retina barrier breakdown in mice and rats

    Journal: Diabetologia

    doi: 10.1007/s00125-013-2998-6

    Overexpression of proNGF induced glial Müller activation and inflammation. Immunohistochemical analysis of retinas was performed 4 weeks after intravitreal injections of pGFP alone (control) ( a–e ), proNGF plus scrambled p75 NTR shRNA (proNGF+Scr)
    Figure Legend Snippet: Overexpression of proNGF induced glial Müller activation and inflammation. Immunohistochemical analysis of retinas was performed 4 weeks after intravitreal injections of pGFP alone (control) ( a–e ), proNGF plus scrambled p75 NTR shRNA (proNGF+Scr)

    Techniques Used: Over Expression, Activation Assay, Immunohistochemistry, shRNA

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    Alomone Labs cleavage resistant prongf
    <t>Overexpression</t> of <t>proNGF</t> induced glial Müller activation and inflammation. Immunohistochemical analysis of retinas was performed 4 weeks after intravitreal injections of pGFP alone (control) ( a–e ), proNGF plus scrambled p75 NTR shRNA (proNGF+Scr)
    Cleavage Resistant Prongf, 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/cleavage resistant prongf/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cleavage resistant prongf - by Bioz Stars, 2022-09
    94/100 stars
      Buy from Supplier

    93
    Alomone Labs pro ngf
    Effect of p75NTR stimulation on lipid-associated genes in Huh7 cells. Huh7 cells were stimulated with 100 ng/ml <t>NGF</t> or 20 ng/ml <t>pro-NGF</t> for 48 h, and the expression of lipid-associated genes was analyzed by qPCR. Primers are shown in Table 2 . Results were normalized to GAPDH and controls set to 1. Values are mean ± SD, n = 3-4. *** p
    Pro Ngf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Overexpression of proNGF induced glial Müller activation and inflammation. Immunohistochemical analysis of retinas was performed 4 weeks after intravitreal injections of pGFP alone (control) ( a–e ), proNGF plus scrambled p75 NTR shRNA (proNGF+Scr)

    Journal: Diabetologia

    Article Title: Modulation of p75NTR prevents diabetes- and proNGF-induced retinal inflammation and blood–retina barrier breakdown in mice and rats

    doi: 10.1007/s00125-013-2998-6

    Figure Lengend Snippet: Overexpression of proNGF induced glial Müller activation and inflammation. Immunohistochemical analysis of retinas was performed 4 weeks after intravitreal injections of pGFP alone (control) ( a–e ), proNGF plus scrambled p75 NTR shRNA (proNGF+Scr)

    Article Snippet: Representative blots ( ) and statistical analysis show that overexpression of cleavage-resistant proNGF plus scrambled shRNA induced a 2.5-fold increase in proNGF , a 1.8-fold increase in p75 , a 3.6-fold increase in NFκB , a 1.7-fold increase in TNF-α ( ) and a 2.3-fold increase in IL-1β ( ).

    Techniques: Over Expression, Activation Assay, Immunohistochemistry, shRNA

    ProNGF activates JNK and NF-κB in primary VS cultures. A . Western blots of protein lysates from primary VS cultures treated Ad5.empty vector (Ad.EV) or Ad5.merlin (Ad.merlin) and probed with anti-phosphorylated JNK antibodies. The blots were stripped

    Journal: Glia

    Article Title: p75NTR is highly expressed in vestibular schwannomas and promotes cell survival by activating NF-κB

    doi: 10.1002/glia.22709

    Figure Lengend Snippet: ProNGF activates JNK and NF-κB in primary VS cultures. A . Western blots of protein lysates from primary VS cultures treated Ad5.empty vector (Ad.EV) or Ad5.merlin (Ad.merlin) and probed with anti-phosphorylated JNK antibodies. The blots were stripped

    Article Snippet: Primary human VS cultures and mouse sciatic nerve Schwann cells maintained in the presence or absence I-JIP or SP600125 (JNK inhibitors) were treated with cleavage resistant proNGF (Alomone Labs, Jerusalem, Israel).

    Techniques: Western Blot, Plasmid Preparation

    ProNGF protects VS cells from apoptosis due to JNK inhibition. A-D . Primary VS cultures were maintained in the presence of the JNK inhibitors, I-JIP or SP600125, with or without treatment with proNGF (3 nM) (A) of proBDNF (3 nM) (D). Cultures were immunostained

    Journal: Glia

    Article Title: p75NTR is highly expressed in vestibular schwannomas and promotes cell survival by activating NF-κB

    doi: 10.1002/glia.22709

    Figure Lengend Snippet: ProNGF protects VS cells from apoptosis due to JNK inhibition. A-D . Primary VS cultures were maintained in the presence of the JNK inhibitors, I-JIP or SP600125, with or without treatment with proNGF (3 nM) (A) of proBDNF (3 nM) (D). Cultures were immunostained

    Article Snippet: Primary human VS cultures and mouse sciatic nerve Schwann cells maintained in the presence or absence I-JIP or SP600125 (JNK inhibitors) were treated with cleavage resistant proNGF (Alomone Labs, Jerusalem, Israel).

    Techniques: Inhibition

    Effect of p75NTR stimulation on lipid-associated genes in Huh7 cells. Huh7 cells were stimulated with 100 ng/ml NGF or 20 ng/ml pro-NGF for 48 h, and the expression of lipid-associated genes was analyzed by qPCR. Primers are shown in Table 2 . Results were normalized to GAPDH and controls set to 1. Values are mean ± SD, n = 3-4. *** p

    Journal: Cell Death & Disease

    Article Title: Caspase-2 and p75 neurotrophin receptor (p75NTR) are involved in the regulation of SREBP and lipid genes in hepatocyte cells

    doi: 10.1038/s41419-019-1758-z

    Figure Lengend Snippet: Effect of p75NTR stimulation on lipid-associated genes in Huh7 cells. Huh7 cells were stimulated with 100 ng/ml NGF or 20 ng/ml pro-NGF for 48 h, and the expression of lipid-associated genes was analyzed by qPCR. Primers are shown in Table 2 . Results were normalized to GAPDH and controls set to 1. Values are mean ± SD, n = 3-4. *** p

    Article Snippet: Cells were stimulated with different concentrations of NGF or pro-NGF (cleavage-resistant, mutant protein) (Alamone Labs) for various periods of times.

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Schematic view of the role of caspase-2 in p75NTR signaling and lipid metabolism. We propose the following scheme for the action of p75NTR and of caspase-2 in SREBP2 and associated gene regulation. Stimulation of p75NTR by NGF or pro-NGF activates the signaling protein p38 MAPK that can phosphorylate caspase-2 (CASP2) leading to the release of caspase-3 (CASP3) from its complex. CASP3, in turn, can cleave SREBP molecules to yield an active transcription factor. SREBP responsive genes include LDLR and other lipogenic genes but also CASP2 itself. Normally SREBP is processed by the enzymes S1P and S2P via a feedback inhibition determined by the cell cholesterol level. We hypothesize that the steroid-mediated and neurotrophin/p75NTR-induced SREBP pathways are distinct but functionally linked

    Journal: Cell Death & Disease

    Article Title: Caspase-2 and p75 neurotrophin receptor (p75NTR) are involved in the regulation of SREBP and lipid genes in hepatocyte cells

    doi: 10.1038/s41419-019-1758-z

    Figure Lengend Snippet: Schematic view of the role of caspase-2 in p75NTR signaling and lipid metabolism. We propose the following scheme for the action of p75NTR and of caspase-2 in SREBP2 and associated gene regulation. Stimulation of p75NTR by NGF or pro-NGF activates the signaling protein p38 MAPK that can phosphorylate caspase-2 (CASP2) leading to the release of caspase-3 (CASP3) from its complex. CASP3, in turn, can cleave SREBP molecules to yield an active transcription factor. SREBP responsive genes include LDLR and other lipogenic genes but also CASP2 itself. Normally SREBP is processed by the enzymes S1P and S2P via a feedback inhibition determined by the cell cholesterol level. We hypothesize that the steroid-mediated and neurotrophin/p75NTR-induced SREBP pathways are distinct but functionally linked

    Article Snippet: Cells were stimulated with different concentrations of NGF or pro-NGF (cleavage-resistant, mutant protein) (Alamone Labs) for various periods of times.

    Techniques: Inhibition

    NGF is expressed in fatty liver of obese mice together with p75NTR. a NGF has increased in leptin-deficient ob/ob mouse livers compared with control, wild-type (wt) mice. b p75NTR was expressed in liver with no significant change between ob/ob and control mice. c , d The level of NGF has increased in leptin-receptor-deficient db/db mouse liver in comparison to controls, c immunoblot analysis and d quantification. ** p

    Journal: Cell Death & Disease

    Article Title: Caspase-2 and p75 neurotrophin receptor (p75NTR) are involved in the regulation of SREBP and lipid genes in hepatocyte cells

    doi: 10.1038/s41419-019-1758-z

    Figure Lengend Snippet: NGF is expressed in fatty liver of obese mice together with p75NTR. a NGF has increased in leptin-deficient ob/ob mouse livers compared with control, wild-type (wt) mice. b p75NTR was expressed in liver with no significant change between ob/ob and control mice. c , d The level of NGF has increased in leptin-receptor-deficient db/db mouse liver in comparison to controls, c immunoblot analysis and d quantification. ** p

    Article Snippet: Cells were stimulated with different concentrations of NGF or pro-NGF (cleavage-resistant, mutant protein) (Alamone Labs) for various periods of times.

    Techniques: Mouse Assay

    p75NTR stimulation activated SREBP1 and SREBP2 in Huh7 hepatocyte cells. Human Huh7 cells were stimulated with 10 ng/ml pro-NGF or 50 ng/ml NGF for 16 h followed by immunoblotting using antibodies to detect the presence of cleaved/activated SREBP1 and SREBP2. a SREBP1, immunoblots and b quantification. Values are mean ± SD, n = 3. ** P

    Journal: Cell Death & Disease

    Article Title: Caspase-2 and p75 neurotrophin receptor (p75NTR) are involved in the regulation of SREBP and lipid genes in hepatocyte cells

    doi: 10.1038/s41419-019-1758-z

    Figure Lengend Snippet: p75NTR stimulation activated SREBP1 and SREBP2 in Huh7 hepatocyte cells. Human Huh7 cells were stimulated with 10 ng/ml pro-NGF or 50 ng/ml NGF for 16 h followed by immunoblotting using antibodies to detect the presence of cleaved/activated SREBP1 and SREBP2. a SREBP1, immunoblots and b quantification. Values are mean ± SD, n = 3. ** P

    Article Snippet: Cells were stimulated with different concentrations of NGF or pro-NGF (cleavage-resistant, mutant protein) (Alamone Labs) for various periods of times.

    Techniques: Western Blot

    Role of Caspase-2 in p75NTR mediated regulation of SREBP. a Structure of human caspase-2 (CASP2) showing the CARD domain and the 18 and 13 kDa caspase subunits. p38 MAPK phosphorylation site at threonine (Thr)180 in CASP2 is conserved among species. The amino acid numbering underlying functional domains in CASP2 is shown. Boxes represent sequence homology among mammalian species. b Phosphoprotein retardation was performed as described in Materials and methods. A specific phospho-CASP2 antibody against Thr180 was used in conjunction with an antibody against CASP2, recognizing non-phosphorylated form of the enzyme. Phosphorylated CASP2 migrated slower in the Phos-Tag gel (right) as compared with the protein run on denaturating SDS-PAGE (left). c , d Huh7 hepatocyte cells were stimulated with 50 ng NGF ( c ) or 10 ng/ml pro-NGF ( d ) for different times followed by immunoblotting using phospho-CASP2 and anti-CASP2 antibodies. β-Actin was used as a control. e , f Cells were treated with pro-NGF for 6 h in the absence or presence 1 μM of the p38 MAPK inhibitor, SB203580 (SB). e Immunoblot and f quantification of p-CASP2 levels. SB reduced the increase in p-CASP2 by pro-NGF. Values are means ± SD, n = 4. * p

    Journal: Cell Death & Disease

    Article Title: Caspase-2 and p75 neurotrophin receptor (p75NTR) are involved in the regulation of SREBP and lipid genes in hepatocyte cells

    doi: 10.1038/s41419-019-1758-z

    Figure Lengend Snippet: Role of Caspase-2 in p75NTR mediated regulation of SREBP. a Structure of human caspase-2 (CASP2) showing the CARD domain and the 18 and 13 kDa caspase subunits. p38 MAPK phosphorylation site at threonine (Thr)180 in CASP2 is conserved among species. The amino acid numbering underlying functional domains in CASP2 is shown. Boxes represent sequence homology among mammalian species. b Phosphoprotein retardation was performed as described in Materials and methods. A specific phospho-CASP2 antibody against Thr180 was used in conjunction with an antibody against CASP2, recognizing non-phosphorylated form of the enzyme. Phosphorylated CASP2 migrated slower in the Phos-Tag gel (right) as compared with the protein run on denaturating SDS-PAGE (left). c , d Huh7 hepatocyte cells were stimulated with 50 ng NGF ( c ) or 10 ng/ml pro-NGF ( d ) for different times followed by immunoblotting using phospho-CASP2 and anti-CASP2 antibodies. β-Actin was used as a control. e , f Cells were treated with pro-NGF for 6 h in the absence or presence 1 μM of the p38 MAPK inhibitor, SB203580 (SB). e Immunoblot and f quantification of p-CASP2 levels. SB reduced the increase in p-CASP2 by pro-NGF. Values are means ± SD, n = 4. * p

    Article Snippet: Cells were stimulated with different concentrations of NGF or pro-NGF (cleavage-resistant, mutant protein) (Alamone Labs) for various periods of times.

    Techniques: Functional Assay, Sequencing, SDS Page