fitc labeled antihuman p2x7r antibody  (Alomone Labs)


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    Name:
    Anti P2X7 Receptor extracellular FITC Antibody
    Description:
    Anti P2X7 Receptor extracellular Antibody APR 008 is a highly specific antibody directed against an epitope of the mouse protein The antibody can be used in western blot indirect flow cytometry immunohistochemistry live cell imaging and immunocytochemistry applications It has been designed to recognize P2X7 purinergic receptor from mouse rat and human samples nAnti P2X7 Receptor extracellular FITC Antibody APR 008 F is directly conjugated to fluorescein isothiocyanate FITC This labeled antibody can be used in immunofluorescent applications such as direct live cell flow cytometry and immunocytochemistry
    Catalog Number:
    APR-008-F
    Price:
    686.0
    Category:
    Primary Antibody
    Applications:
    Flow Cytometry, Immunocytochemistry, Immunofluorescence, Live Cell Imaging
    Purity:
    Affinity purified on immobilized antigen.
    Immunogen:
    Synthetic peptide
    Size:
    50 mcl
    Antibody Type:
    Polyclonal FITC Conjugated Primary Antibody
    Format:
    Lyophilized Powder
    Host:
    Rabbit
    Isotype:
    Rabbit IgG
    Buy from Supplier


    Structured Review

    Alomone Labs fitc labeled antihuman p2x7r antibody
    Anti P2X7 Receptor extracellular FITC Antibody
    Anti P2X7 Receptor extracellular Antibody APR 008 is a highly specific antibody directed against an epitope of the mouse protein The antibody can be used in western blot indirect flow cytometry immunohistochemistry live cell imaging and immunocytochemistry applications It has been designed to recognize P2X7 purinergic receptor from mouse rat and human samples nAnti P2X7 Receptor extracellular FITC Antibody APR 008 F is directly conjugated to fluorescein isothiocyanate FITC This labeled antibody can be used in immunofluorescent applications such as direct live cell flow cytometry and immunocytochemistry
    https://www.bioz.com/result/fitc labeled antihuman p2x7r antibody/product/Alomone Labs
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    fitc labeled antihuman p2x7r antibody - by Bioz Stars, 2021-09
    92/100 stars

    Images

    1) Product Images from "Differential expression of P2X7 receptor and IL-1β in nociceptive and neuropathic pain"

    Article Title: Differential expression of P2X7 receptor and IL-1β in nociceptive and neuropathic pain

    Journal: Journal of Neuroinflammation

    doi: 10.1186/s12974-016-0565-z

    P2X7R expression on lymphocytes ( a ), CD4 + ( b ) and CD4 − cells ( c ), and monocytes ( d ) is elevated in patients with neuropathic pain. PBMCs were stained with PerCP-labeled antihuman CD4 antibody and FITC-labeled antihuman P2X7R antibody and analyzed by FACS. The results show a significantly elevated P2X7R expression only in patients suffering from neuropathic pain on the analyzed cell type (lymphocytes ( p = 0.009), CD4 + cells ( p
    Figure Legend Snippet: P2X7R expression on lymphocytes ( a ), CD4 + ( b ) and CD4 − cells ( c ), and monocytes ( d ) is elevated in patients with neuropathic pain. PBMCs were stained with PerCP-labeled antihuman CD4 antibody and FITC-labeled antihuman P2X7R antibody and analyzed by FACS. The results show a significantly elevated P2X7R expression only in patients suffering from neuropathic pain on the analyzed cell type (lymphocytes ( p = 0.009), CD4 + cells ( p

    Techniques Used: Expressing, Staining, Labeling, FACS

    IL-1β levels are elevated in patients with neuropathic pain. As IL-1β was shown to be the key mediator in P2X7R-pain interplay, we analyzed serum levels of this pro-inflammatory and pro-nociceptive cytokine by multiplex immune assay. Affirmatively to the results with elevated P2X7Rs, we solely found significant increased serum levels in the peripheral blood of patients with NeP * p
    Figure Legend Snippet: IL-1β levels are elevated in patients with neuropathic pain. As IL-1β was shown to be the key mediator in P2X7R-pain interplay, we analyzed serum levels of this pro-inflammatory and pro-nociceptive cytokine by multiplex immune assay. Affirmatively to the results with elevated P2X7Rs, we solely found significant increased serum levels in the peripheral blood of patients with NeP * p

    Techniques Used: Multiplex Assay

    Exemplary illustration of P2X7 receptor expression. For quantitative analysis of the proportion of CD4 + cells ( a ) and P2X7R protein expression ( b ), cells were stained with PerCP-labeled antihuman CD4 antibody and FITC-labeled antihuman P2X7R antibody. We separately analyzed each subgroup by the mean fluorescence intensity (MFI) of P2X7R expression. Overlay histograms of representative results of P2X7R expression of one patient/healthy volunteer are displayed in histograms
    Figure Legend Snippet: Exemplary illustration of P2X7 receptor expression. For quantitative analysis of the proportion of CD4 + cells ( a ) and P2X7R protein expression ( b ), cells were stained with PerCP-labeled antihuman CD4 antibody and FITC-labeled antihuman P2X7R antibody. We separately analyzed each subgroup by the mean fluorescence intensity (MFI) of P2X7R expression. Overlay histograms of representative results of P2X7R expression of one patient/healthy volunteer are displayed in histograms

    Techniques Used: Expressing, Staining, Labeling, Fluorescence

    P2RX7 mRNA expression is exclusively increased in patients with neuropathic pain. To confirm the flow cytometric observations with elevated P2X7R protein expression, predominantly on CD4 + cells and only in NeP, we determined the relative P2RX7 mRNA expression of CD4 + cells by qPCR. Affirmatively, increased mRNA levels were consistent with the flow cytometric analyses and exclusively elevated in patients with NeP. ( p = 0.038). * p
    Figure Legend Snippet: P2RX7 mRNA expression is exclusively increased in patients with neuropathic pain. To confirm the flow cytometric observations with elevated P2X7R protein expression, predominantly on CD4 + cells and only in NeP, we determined the relative P2RX7 mRNA expression of CD4 + cells by qPCR. Affirmatively, increased mRNA levels were consistent with the flow cytometric analyses and exclusively elevated in patients with NeP. ( p = 0.038). * p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    Differential blood count and quantification of CD4 + cells. In order to avoid misinterpretation of potentially elevated P2X7R protein expression based on different cell counts, we quantified numbers of polymorphonuclear leukocytes ( a ), lymphocytes ( b ), and CD4 + cells ( c ). No differences were found between patients suffering from CLBP, NeP, and healthy volunteers. Data are presented as mean ± SD, n = 19, * p
    Figure Legend Snippet: Differential blood count and quantification of CD4 + cells. In order to avoid misinterpretation of potentially elevated P2X7R protein expression based on different cell counts, we quantified numbers of polymorphonuclear leukocytes ( a ), lymphocytes ( b ), and CD4 + cells ( c ). No differences were found between patients suffering from CLBP, NeP, and healthy volunteers. Data are presented as mean ± SD, n = 19, * p

    Techniques Used: Expressing

    2) Product Images from "P2X7R mutation disrupts the NLRP3-mediated Th program and predicts poor cardiac allograft outcomes"

    Article Title: P2X7R mutation disrupts the NLRP3-mediated Th program and predicts poor cardiac allograft outcomes

    Journal: The Journal of Clinical Investigation

    doi: 10.1172/JCI94524

    A C-terminal P2X7R mutation is associated with poor outcomes in cardiac-transplanted patients. ( A ) Bar graph depicting the percentage of cardiac-transplanted patients who carry the WT or mutant P2X7R allele ( n = 102) with an MIT change greater than 0.5 mm (early cardiac allograft vasculopathy, in black) at 1 year after transplantation in the CTOT-05 cohort. ( B ) Bar graph depicting the number of acute rejection episodes in cardiac-transplanted patients who carry the WT (black) or mutant (white) P2X7R allele ( n = 181) within the first year after transplant in the NIT -Bergamo cohort. ( C ) Bar graph depicting the percentage of cardiac-transplanted patients who carry the WT or mutant P2X7R allele ( n = 130) with major adverse cardiac events (MACEs, in black) at 10 years of follow-up in the AIRT -Bologna cohort. In A and C : black, percentage of patients who experienced the event; white, percentage who were free from events. ( D ) Line graph depicting the estimated odds ratio (OR) for clinical outcomes recorded in the 3 cohorts of cardiac-transplanted patients who carry the WT or mutant P2X7R allele. In the NIT -Bergamo cohort, the OR was calculated based on the requirement of medical intervention for acute rejection episodes with a frequency of greater or less than 3 episodes. * P
    Figure Legend Snippet: A C-terminal P2X7R mutation is associated with poor outcomes in cardiac-transplanted patients. ( A ) Bar graph depicting the percentage of cardiac-transplanted patients who carry the WT or mutant P2X7R allele ( n = 102) with an MIT change greater than 0.5 mm (early cardiac allograft vasculopathy, in black) at 1 year after transplantation in the CTOT-05 cohort. ( B ) Bar graph depicting the number of acute rejection episodes in cardiac-transplanted patients who carry the WT (black) or mutant (white) P2X7R allele ( n = 181) within the first year after transplant in the NIT -Bergamo cohort. ( C ) Bar graph depicting the percentage of cardiac-transplanted patients who carry the WT or mutant P2X7R allele ( n = 130) with major adverse cardiac events (MACEs, in black) at 10 years of follow-up in the AIRT -Bologna cohort. In A and C : black, percentage of patients who experienced the event; white, percentage who were free from events. ( D ) Line graph depicting the estimated odds ratio (OR) for clinical outcomes recorded in the 3 cohorts of cardiac-transplanted patients who carry the WT or mutant P2X7R allele. In the NIT -Bergamo cohort, the OR was calculated based on the requirement of medical intervention for acute rejection episodes with a frequency of greater or less than 3 episodes. * P

    Techniques Used: Mutagenesis, Transplantation Assay

    P2X7R loss of function in a preclinical model of heart transplantation reduces allograft survival, with increased vasculopathy and Th1/Th17–mediated responses. ( A ) P2X7R –/– mice receiving bm12 heart transplantation demonstrated reduced graft survival as compared with B6 recipients (** P
    Figure Legend Snippet: P2X7R loss of function in a preclinical model of heart transplantation reduces allograft survival, with increased vasculopathy and Th1/Th17–mediated responses. ( A ) P2X7R –/– mice receiving bm12 heart transplantation demonstrated reduced graft survival as compared with B6 recipients (** P

    Techniques Used: Transplantation Assay, Mouse Assay

    The dysregulated P2X7R/NLRP3 pathway is associated with altered Th cell fate.
    Figure Legend Snippet: The dysregulated P2X7R/NLRP3 pathway is associated with altered Th cell fate.

    Techniques Used:

    Existence of a P2X7R/NLRP3 pathway within human CD4 + T cells. ( A ) P2X7R and NLRP3 immunoprecipitation (IP) in human CD4 + T cells. Expression of NLRP3 (top blot) and P2X7R (bottom blot) is shown. Lane 1: Total protein. Lane 2: IP with NLRP3 Ab. Lane 3: IP with P2X7R Ab. Lane 4: IP with Ab alone (NLRP3 and P2X7R). Lane 5: IP with control IgG (for NLRP3 Ab in top blot, for P2X7R Ab in bottom blot). The experiment was run in triplicate (representative blot shown). ( B and C ) Confocal microscopy analysis ( B , scale bar: 5 μm, ×100 original magnification; C , scale bars: 20 μm, ×40 original magnification) depicting baseline colocalization of P2X7R (green) and NLRP3 (red) in human CD4 + T cells. Cells were stained with DAPI (blue) and immunolabeled with anti-P2X7R (green) and anti-NLRP3 Abs (red) ( n = 3). ( D – F ) Bar graphs depicting expression of NLRP3 mRNA by qRT-PCR ( D ), and protein by flow cytometry ( E ) and ELISA ( F ), evaluated in human CD4 + T cells activated with benzoyl ATP (BzATP) and treated with CE-224,535, a P2X7R inhibitor. Experiments were run in duplicate ( n = 5). ( G ) Bar graph representing expression of NLRP3 on human CD4 + P2X7R + cells analyzed by flow cytometry upon BzATP stimulation ( n = 5). ( H ) Representative flow dot plots of NLRP3 expression upon gating on human BzATP-stimulated CD4 + P2X7R + cells. ( I ) Confocal analysis (scale bar: 5 μm; ×100 original magnification) depicting colocalization of P2X7R (green) and NLRP3 (red) in CD4 + T cells upon in vitro stimulation of P2X7R with BzATP ( n = 3). ( J – M ) Bar graphs comparing expression of NLRP3 downstream signaling Th2-related factors IL-4 ( J ), IRF4 ( K ), GATA-3 ( L ), and IL-10 ( M ) by qRT-PCR using mRNA isolated from human CD4 + T cells activated with BzATP and treated with the P2X7R inhibitor CE-224,535. Experiments were run in triplicate ( n = 5). Data are expressed as mean ± SEM. * P
    Figure Legend Snippet: Existence of a P2X7R/NLRP3 pathway within human CD4 + T cells. ( A ) P2X7R and NLRP3 immunoprecipitation (IP) in human CD4 + T cells. Expression of NLRP3 (top blot) and P2X7R (bottom blot) is shown. Lane 1: Total protein. Lane 2: IP with NLRP3 Ab. Lane 3: IP with P2X7R Ab. Lane 4: IP with Ab alone (NLRP3 and P2X7R). Lane 5: IP with control IgG (for NLRP3 Ab in top blot, for P2X7R Ab in bottom blot). The experiment was run in triplicate (representative blot shown). ( B and C ) Confocal microscopy analysis ( B , scale bar: 5 μm, ×100 original magnification; C , scale bars: 20 μm, ×40 original magnification) depicting baseline colocalization of P2X7R (green) and NLRP3 (red) in human CD4 + T cells. Cells were stained with DAPI (blue) and immunolabeled with anti-P2X7R (green) and anti-NLRP3 Abs (red) ( n = 3). ( D – F ) Bar graphs depicting expression of NLRP3 mRNA by qRT-PCR ( D ), and protein by flow cytometry ( E ) and ELISA ( F ), evaluated in human CD4 + T cells activated with benzoyl ATP (BzATP) and treated with CE-224,535, a P2X7R inhibitor. Experiments were run in duplicate ( n = 5). ( G ) Bar graph representing expression of NLRP3 on human CD4 + P2X7R + cells analyzed by flow cytometry upon BzATP stimulation ( n = 5). ( H ) Representative flow dot plots of NLRP3 expression upon gating on human BzATP-stimulated CD4 + P2X7R + cells. ( I ) Confocal analysis (scale bar: 5 μm; ×100 original magnification) depicting colocalization of P2X7R (green) and NLRP3 (red) in CD4 + T cells upon in vitro stimulation of P2X7R with BzATP ( n = 3). ( J – M ) Bar graphs comparing expression of NLRP3 downstream signaling Th2-related factors IL-4 ( J ), IRF4 ( K ), GATA-3 ( L ), and IL-10 ( M ) by qRT-PCR using mRNA isolated from human CD4 + T cells activated with BzATP and treated with the P2X7R inhibitor CE-224,535. Experiments were run in triplicate ( n = 5). Data are expressed as mean ± SEM. * P

    Techniques Used: Immunoprecipitation, Expressing, Confocal Microscopy, Staining, Immunolabeling, Quantitative RT-PCR, Flow Cytometry, Enzyme-linked Immunosorbent Assay, In Vitro, Isolation

    Related Articles

    Staining:

    Article Title: Differential expression of P2X7 receptor and IL-1β in nociceptive and neuropathic pain
    Article Snippet: .. For extracellular staining, cells were co-incubated with PerCP-labeled antihuman CD4 antibody (1:50, Biolegend, San Diego, CA, USA) and FITC-labeled antihuman P2X7R antibody (1:100, Alomone Labs, Jerusalem, Israel) at room temperature for 1 hour. ..

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    Alomone Labs fitc labeled antihuman p2x7r antibody
    <t>P2X7R</t> expression on lymphocytes ( a ), CD4 + ( b ) and CD4 − cells ( c ), and monocytes ( d ) is elevated in patients with neuropathic pain. PBMCs were stained with PerCP-labeled antihuman CD4 antibody and FITC-labeled antihuman P2X7R antibody and analyzed by FACS. The results show a significantly elevated P2X7R expression only in patients suffering from neuropathic pain on the analyzed cell type (lymphocytes ( p = 0.009), CD4 + cells ( p
    Fitc Labeled Antihuman P2x7r Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fitc labeled antihuman p2x7r antibody/product/Alomone Labs
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    fitc labeled antihuman p2x7r antibody - by Bioz Stars, 2021-09
    92/100 stars
      Buy from Supplier

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    P2X7R expression on lymphocytes ( a ), CD4 + ( b ) and CD4 − cells ( c ), and monocytes ( d ) is elevated in patients with neuropathic pain. PBMCs were stained with PerCP-labeled antihuman CD4 antibody and FITC-labeled antihuman P2X7R antibody and analyzed by FACS. The results show a significantly elevated P2X7R expression only in patients suffering from neuropathic pain on the analyzed cell type (lymphocytes ( p = 0.009), CD4 + cells ( p

    Journal: Journal of Neuroinflammation

    Article Title: Differential expression of P2X7 receptor and IL-1β in nociceptive and neuropathic pain

    doi: 10.1186/s12974-016-0565-z

    Figure Lengend Snippet: P2X7R expression on lymphocytes ( a ), CD4 + ( b ) and CD4 − cells ( c ), and monocytes ( d ) is elevated in patients with neuropathic pain. PBMCs were stained with PerCP-labeled antihuman CD4 antibody and FITC-labeled antihuman P2X7R antibody and analyzed by FACS. The results show a significantly elevated P2X7R expression only in patients suffering from neuropathic pain on the analyzed cell type (lymphocytes ( p = 0.009), CD4 + cells ( p

    Article Snippet: For extracellular staining, cells were co-incubated with PerCP-labeled antihuman CD4 antibody (1:50, Biolegend, San Diego, CA, USA) and FITC-labeled antihuman P2X7R antibody (1:100, Alomone Labs, Jerusalem, Israel) at room temperature for 1 hour.

    Techniques: Expressing, Staining, Labeling, FACS

    IL-1β levels are elevated in patients with neuropathic pain. As IL-1β was shown to be the key mediator in P2X7R-pain interplay, we analyzed serum levels of this pro-inflammatory and pro-nociceptive cytokine by multiplex immune assay. Affirmatively to the results with elevated P2X7Rs, we solely found significant increased serum levels in the peripheral blood of patients with NeP * p

    Journal: Journal of Neuroinflammation

    Article Title: Differential expression of P2X7 receptor and IL-1β in nociceptive and neuropathic pain

    doi: 10.1186/s12974-016-0565-z

    Figure Lengend Snippet: IL-1β levels are elevated in patients with neuropathic pain. As IL-1β was shown to be the key mediator in P2X7R-pain interplay, we analyzed serum levels of this pro-inflammatory and pro-nociceptive cytokine by multiplex immune assay. Affirmatively to the results with elevated P2X7Rs, we solely found significant increased serum levels in the peripheral blood of patients with NeP * p

    Article Snippet: For extracellular staining, cells were co-incubated with PerCP-labeled antihuman CD4 antibody (1:50, Biolegend, San Diego, CA, USA) and FITC-labeled antihuman P2X7R antibody (1:100, Alomone Labs, Jerusalem, Israel) at room temperature for 1 hour.

    Techniques: Multiplex Assay

    Exemplary illustration of P2X7 receptor expression. For quantitative analysis of the proportion of CD4 + cells ( a ) and P2X7R protein expression ( b ), cells were stained with PerCP-labeled antihuman CD4 antibody and FITC-labeled antihuman P2X7R antibody. We separately analyzed each subgroup by the mean fluorescence intensity (MFI) of P2X7R expression. Overlay histograms of representative results of P2X7R expression of one patient/healthy volunteer are displayed in histograms

    Journal: Journal of Neuroinflammation

    Article Title: Differential expression of P2X7 receptor and IL-1β in nociceptive and neuropathic pain

    doi: 10.1186/s12974-016-0565-z

    Figure Lengend Snippet: Exemplary illustration of P2X7 receptor expression. For quantitative analysis of the proportion of CD4 + cells ( a ) and P2X7R protein expression ( b ), cells were stained with PerCP-labeled antihuman CD4 antibody and FITC-labeled antihuman P2X7R antibody. We separately analyzed each subgroup by the mean fluorescence intensity (MFI) of P2X7R expression. Overlay histograms of representative results of P2X7R expression of one patient/healthy volunteer are displayed in histograms

    Article Snippet: For extracellular staining, cells were co-incubated with PerCP-labeled antihuman CD4 antibody (1:50, Biolegend, San Diego, CA, USA) and FITC-labeled antihuman P2X7R antibody (1:100, Alomone Labs, Jerusalem, Israel) at room temperature for 1 hour.

    Techniques: Expressing, Staining, Labeling, Fluorescence

    P2RX7 mRNA expression is exclusively increased in patients with neuropathic pain. To confirm the flow cytometric observations with elevated P2X7R protein expression, predominantly on CD4 + cells and only in NeP, we determined the relative P2RX7 mRNA expression of CD4 + cells by qPCR. Affirmatively, increased mRNA levels were consistent with the flow cytometric analyses and exclusively elevated in patients with NeP. ( p = 0.038). * p

    Journal: Journal of Neuroinflammation

    Article Title: Differential expression of P2X7 receptor and IL-1β in nociceptive and neuropathic pain

    doi: 10.1186/s12974-016-0565-z

    Figure Lengend Snippet: P2RX7 mRNA expression is exclusively increased in patients with neuropathic pain. To confirm the flow cytometric observations with elevated P2X7R protein expression, predominantly on CD4 + cells and only in NeP, we determined the relative P2RX7 mRNA expression of CD4 + cells by qPCR. Affirmatively, increased mRNA levels were consistent with the flow cytometric analyses and exclusively elevated in patients with NeP. ( p = 0.038). * p

    Article Snippet: For extracellular staining, cells were co-incubated with PerCP-labeled antihuman CD4 antibody (1:50, Biolegend, San Diego, CA, USA) and FITC-labeled antihuman P2X7R antibody (1:100, Alomone Labs, Jerusalem, Israel) at room temperature for 1 hour.

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Differential blood count and quantification of CD4 + cells. In order to avoid misinterpretation of potentially elevated P2X7R protein expression based on different cell counts, we quantified numbers of polymorphonuclear leukocytes ( a ), lymphocytes ( b ), and CD4 + cells ( c ). No differences were found between patients suffering from CLBP, NeP, and healthy volunteers. Data are presented as mean ± SD, n = 19, * p

    Journal: Journal of Neuroinflammation

    Article Title: Differential expression of P2X7 receptor and IL-1β in nociceptive and neuropathic pain

    doi: 10.1186/s12974-016-0565-z

    Figure Lengend Snippet: Differential blood count and quantification of CD4 + cells. In order to avoid misinterpretation of potentially elevated P2X7R protein expression based on different cell counts, we quantified numbers of polymorphonuclear leukocytes ( a ), lymphocytes ( b ), and CD4 + cells ( c ). No differences were found between patients suffering from CLBP, NeP, and healthy volunteers. Data are presented as mean ± SD, n = 19, * p

    Article Snippet: For extracellular staining, cells were co-incubated with PerCP-labeled antihuman CD4 antibody (1:50, Biolegend, San Diego, CA, USA) and FITC-labeled antihuman P2X7R antibody (1:100, Alomone Labs, Jerusalem, Israel) at room temperature for 1 hour.

    Techniques: Expressing

    A C-terminal P2X7R mutation is associated with poor outcomes in cardiac-transplanted patients. ( A ) Bar graph depicting the percentage of cardiac-transplanted patients who carry the WT or mutant P2X7R allele ( n = 102) with an MIT change greater than 0.5 mm (early cardiac allograft vasculopathy, in black) at 1 year after transplantation in the CTOT-05 cohort. ( B ) Bar graph depicting the number of acute rejection episodes in cardiac-transplanted patients who carry the WT (black) or mutant (white) P2X7R allele ( n = 181) within the first year after transplant in the NIT -Bergamo cohort. ( C ) Bar graph depicting the percentage of cardiac-transplanted patients who carry the WT or mutant P2X7R allele ( n = 130) with major adverse cardiac events (MACEs, in black) at 10 years of follow-up in the AIRT -Bologna cohort. In A and C : black, percentage of patients who experienced the event; white, percentage who were free from events. ( D ) Line graph depicting the estimated odds ratio (OR) for clinical outcomes recorded in the 3 cohorts of cardiac-transplanted patients who carry the WT or mutant P2X7R allele. In the NIT -Bergamo cohort, the OR was calculated based on the requirement of medical intervention for acute rejection episodes with a frequency of greater or less than 3 episodes. * P

    Journal: The Journal of Clinical Investigation

    Article Title: P2X7R mutation disrupts the NLRP3-mediated Th program and predicts poor cardiac allograft outcomes

    doi: 10.1172/JCI94524

    Figure Lengend Snippet: A C-terminal P2X7R mutation is associated with poor outcomes in cardiac-transplanted patients. ( A ) Bar graph depicting the percentage of cardiac-transplanted patients who carry the WT or mutant P2X7R allele ( n = 102) with an MIT change greater than 0.5 mm (early cardiac allograft vasculopathy, in black) at 1 year after transplantation in the CTOT-05 cohort. ( B ) Bar graph depicting the number of acute rejection episodes in cardiac-transplanted patients who carry the WT (black) or mutant (white) P2X7R allele ( n = 181) within the first year after transplant in the NIT -Bergamo cohort. ( C ) Bar graph depicting the percentage of cardiac-transplanted patients who carry the WT or mutant P2X7R allele ( n = 130) with major adverse cardiac events (MACEs, in black) at 10 years of follow-up in the AIRT -Bologna cohort. In A and C : black, percentage of patients who experienced the event; white, percentage who were free from events. ( D ) Line graph depicting the estimated odds ratio (OR) for clinical outcomes recorded in the 3 cohorts of cardiac-transplanted patients who carry the WT or mutant P2X7R allele. In the NIT -Bergamo cohort, the OR was calculated based on the requirement of medical intervention for acute rejection episodes with a frequency of greater or less than 3 episodes. * P

    Article Snippet: FITC-conjugated anti–mouse P2X7R (APR-008-F) was purchased from Alomone Labs.

    Techniques: Mutagenesis, Transplantation Assay

    P2X7R loss of function in a preclinical model of heart transplantation reduces allograft survival, with increased vasculopathy and Th1/Th17–mediated responses. ( A ) P2X7R –/– mice receiving bm12 heart transplantation demonstrated reduced graft survival as compared with B6 recipients (** P

    Journal: The Journal of Clinical Investigation

    Article Title: P2X7R mutation disrupts the NLRP3-mediated Th program and predicts poor cardiac allograft outcomes

    doi: 10.1172/JCI94524

    Figure Lengend Snippet: P2X7R loss of function in a preclinical model of heart transplantation reduces allograft survival, with increased vasculopathy and Th1/Th17–mediated responses. ( A ) P2X7R –/– mice receiving bm12 heart transplantation demonstrated reduced graft survival as compared with B6 recipients (** P

    Article Snippet: FITC-conjugated anti–mouse P2X7R (APR-008-F) was purchased from Alomone Labs.

    Techniques: Transplantation Assay, Mouse Assay

    The dysregulated P2X7R/NLRP3 pathway is associated with altered Th cell fate.

    Journal: The Journal of Clinical Investigation

    Article Title: P2X7R mutation disrupts the NLRP3-mediated Th program and predicts poor cardiac allograft outcomes

    doi: 10.1172/JCI94524

    Figure Lengend Snippet: The dysregulated P2X7R/NLRP3 pathway is associated with altered Th cell fate.

    Article Snippet: FITC-conjugated anti–mouse P2X7R (APR-008-F) was purchased from Alomone Labs.

    Techniques:

    Existence of a P2X7R/NLRP3 pathway within human CD4 + T cells. ( A ) P2X7R and NLRP3 immunoprecipitation (IP) in human CD4 + T cells. Expression of NLRP3 (top blot) and P2X7R (bottom blot) is shown. Lane 1: Total protein. Lane 2: IP with NLRP3 Ab. Lane 3: IP with P2X7R Ab. Lane 4: IP with Ab alone (NLRP3 and P2X7R). Lane 5: IP with control IgG (for NLRP3 Ab in top blot, for P2X7R Ab in bottom blot). The experiment was run in triplicate (representative blot shown). ( B and C ) Confocal microscopy analysis ( B , scale bar: 5 μm, ×100 original magnification; C , scale bars: 20 μm, ×40 original magnification) depicting baseline colocalization of P2X7R (green) and NLRP3 (red) in human CD4 + T cells. Cells were stained with DAPI (blue) and immunolabeled with anti-P2X7R (green) and anti-NLRP3 Abs (red) ( n = 3). ( D – F ) Bar graphs depicting expression of NLRP3 mRNA by qRT-PCR ( D ), and protein by flow cytometry ( E ) and ELISA ( F ), evaluated in human CD4 + T cells activated with benzoyl ATP (BzATP) and treated with CE-224,535, a P2X7R inhibitor. Experiments were run in duplicate ( n = 5). ( G ) Bar graph representing expression of NLRP3 on human CD4 + P2X7R + cells analyzed by flow cytometry upon BzATP stimulation ( n = 5). ( H ) Representative flow dot plots of NLRP3 expression upon gating on human BzATP-stimulated CD4 + P2X7R + cells. ( I ) Confocal analysis (scale bar: 5 μm; ×100 original magnification) depicting colocalization of P2X7R (green) and NLRP3 (red) in CD4 + T cells upon in vitro stimulation of P2X7R with BzATP ( n = 3). ( J – M ) Bar graphs comparing expression of NLRP3 downstream signaling Th2-related factors IL-4 ( J ), IRF4 ( K ), GATA-3 ( L ), and IL-10 ( M ) by qRT-PCR using mRNA isolated from human CD4 + T cells activated with BzATP and treated with the P2X7R inhibitor CE-224,535. Experiments were run in triplicate ( n = 5). Data are expressed as mean ± SEM. * P

    Journal: The Journal of Clinical Investigation

    Article Title: P2X7R mutation disrupts the NLRP3-mediated Th program and predicts poor cardiac allograft outcomes

    doi: 10.1172/JCI94524

    Figure Lengend Snippet: Existence of a P2X7R/NLRP3 pathway within human CD4 + T cells. ( A ) P2X7R and NLRP3 immunoprecipitation (IP) in human CD4 + T cells. Expression of NLRP3 (top blot) and P2X7R (bottom blot) is shown. Lane 1: Total protein. Lane 2: IP with NLRP3 Ab. Lane 3: IP with P2X7R Ab. Lane 4: IP with Ab alone (NLRP3 and P2X7R). Lane 5: IP with control IgG (for NLRP3 Ab in top blot, for P2X7R Ab in bottom blot). The experiment was run in triplicate (representative blot shown). ( B and C ) Confocal microscopy analysis ( B , scale bar: 5 μm, ×100 original magnification; C , scale bars: 20 μm, ×40 original magnification) depicting baseline colocalization of P2X7R (green) and NLRP3 (red) in human CD4 + T cells. Cells were stained with DAPI (blue) and immunolabeled with anti-P2X7R (green) and anti-NLRP3 Abs (red) ( n = 3). ( D – F ) Bar graphs depicting expression of NLRP3 mRNA by qRT-PCR ( D ), and protein by flow cytometry ( E ) and ELISA ( F ), evaluated in human CD4 + T cells activated with benzoyl ATP (BzATP) and treated with CE-224,535, a P2X7R inhibitor. Experiments were run in duplicate ( n = 5). ( G ) Bar graph representing expression of NLRP3 on human CD4 + P2X7R + cells analyzed by flow cytometry upon BzATP stimulation ( n = 5). ( H ) Representative flow dot plots of NLRP3 expression upon gating on human BzATP-stimulated CD4 + P2X7R + cells. ( I ) Confocal analysis (scale bar: 5 μm; ×100 original magnification) depicting colocalization of P2X7R (green) and NLRP3 (red) in CD4 + T cells upon in vitro stimulation of P2X7R with BzATP ( n = 3). ( J – M ) Bar graphs comparing expression of NLRP3 downstream signaling Th2-related factors IL-4 ( J ), IRF4 ( K ), GATA-3 ( L ), and IL-10 ( M ) by qRT-PCR using mRNA isolated from human CD4 + T cells activated with BzATP and treated with the P2X7R inhibitor CE-224,535. Experiments were run in triplicate ( n = 5). Data are expressed as mean ± SEM. * P

    Article Snippet: FITC-conjugated anti–mouse P2X7R (APR-008-F) was purchased from Alomone Labs.

    Techniques: Immunoprecipitation, Expressing, Confocal Microscopy, Staining, Immunolabeling, Quantitative RT-PCR, Flow Cytometry, Enzyme-linked Immunosorbent Assay, In Vitro, Isolation