fitc labeled antihuman p2x7r antibody  (Alomone Labs)


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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, 2022-01
    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

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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, 2022-01
    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