anti p2x7r  (Alomone Labs)


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    Alomone Labs anti p2x7r
    Primary antibodies used for Western Blot (WB) and immunohistochemistry (IHC).
    Anti P2x7r, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "A Co-Culture System with an Organotypic Lung Slice and an Immortal Alveolar Macrophage Cell Line to Quantify Silica-Induced Inflammation"

    Article Title: A Co-Culture System with an Organotypic Lung Slice and an Immortal Alveolar Macrophage Cell Line to Quantify Silica-Induced Inflammation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0117056

    Primary antibodies used for Western Blot (WB) and immunohistochemistry (IHC).
    Figure Legend Snippet: Primary antibodies used for Western Blot (WB) and immunohistochemistry (IHC).

    Techniques Used: Western Blot, Immunohistochemistry

    p2x7r staining  (Alomone Labs)


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    Alomone Labs p2x7r staining
    <t>P2X7R</t> is present in neurons cultivated in vitro during development. A , Scheme of the experiment. B , Scheme of the regions amplified by qPCR. C , Analysis of the expression of extracellular and ( D ) intracellular sequences of P2X7R by real-time qPCR (DIV1) on each DIV. Relative expression is presented as the fold change normalized to GAPDH expression. E , WT cells were cultured for the indicated period of time, and P2X7R protein expression was determined by immunoblotting. Graphs represent densitometric evaluation of the target protein band normalized to the total amount of loaded protein ( n = 4). F , To validate the specificity of the P2X7R antibody, WT and P2X7R-KO cells were cultured for the indicated period of time, and P2X7R protein expression was determined by immunoblotting. Graphs represent the densitometric evaluation of the target protein band normalized to total amount of loaded protein ( n = 4). G , Areas under the curve for WT and KO primary pyramidal cells at DIV4 after acute BzATP (1 m m ) application. H , Areas under the curve for WT and KO primary pyramidal cells at DIV4 after acute calcium ionophore (1 m m ) application. I , Immunostaining of a neuronal marker (MAP2) and P2X7R in KO and WT primary neurons at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 100 µm. J , Immunostaining for a transfection marker (eGFP) and P2X7R at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 200 µm. * p < 0.05. ** p < 0.005. **** p < 0.0001. ns, non-significant. Data presented as Mean±SEM.
    P2x7r Staining, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 86 stars, based on 1 article reviews
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    p2x7r staining - by Bioz Stars, 2023-03
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    Images

    1) Product Images from "Dual Role of the P2X7 Receptor in Dendritic Outgrowth during Physiological and Pathological Brain Development"

    Article Title: Dual Role of the P2X7 Receptor in Dendritic Outgrowth during Physiological and Pathological Brain Development

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.0805-22.2022

    P2X7R is present in neurons cultivated in vitro during development. A , Scheme of the experiment. B , Scheme of the regions amplified by qPCR. C , Analysis of the expression of extracellular and ( D ) intracellular sequences of P2X7R by real-time qPCR (DIV1) on each DIV. Relative expression is presented as the fold change normalized to GAPDH expression. E , WT cells were cultured for the indicated period of time, and P2X7R protein expression was determined by immunoblotting. Graphs represent densitometric evaluation of the target protein band normalized to the total amount of loaded protein ( n = 4). F , To validate the specificity of the P2X7R antibody, WT and P2X7R-KO cells were cultured for the indicated period of time, and P2X7R protein expression was determined by immunoblotting. Graphs represent the densitometric evaluation of the target protein band normalized to total amount of loaded protein ( n = 4). G , Areas under the curve for WT and KO primary pyramidal cells at DIV4 after acute BzATP (1 m m ) application. H , Areas under the curve for WT and KO primary pyramidal cells at DIV4 after acute calcium ionophore (1 m m ) application. I , Immunostaining of a neuronal marker (MAP2) and P2X7R in KO and WT primary neurons at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 100 µm. J , Immunostaining for a transfection marker (eGFP) and P2X7R at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 200 µm. * p < 0.05. ** p < 0.005. **** p < 0.0001. ns, non-significant. Data presented as Mean±SEM.
    Figure Legend Snippet: P2X7R is present in neurons cultivated in vitro during development. A , Scheme of the experiment. B , Scheme of the regions amplified by qPCR. C , Analysis of the expression of extracellular and ( D ) intracellular sequences of P2X7R by real-time qPCR (DIV1) on each DIV. Relative expression is presented as the fold change normalized to GAPDH expression. E , WT cells were cultured for the indicated period of time, and P2X7R protein expression was determined by immunoblotting. Graphs represent densitometric evaluation of the target protein band normalized to the total amount of loaded protein ( n = 4). F , To validate the specificity of the P2X7R antibody, WT and P2X7R-KO cells were cultured for the indicated period of time, and P2X7R protein expression was determined by immunoblotting. Graphs represent the densitometric evaluation of the target protein band normalized to total amount of loaded protein ( n = 4). G , Areas under the curve for WT and KO primary pyramidal cells at DIV4 after acute BzATP (1 m m ) application. H , Areas under the curve for WT and KO primary pyramidal cells at DIV4 after acute calcium ionophore (1 m m ) application. I , Immunostaining of a neuronal marker (MAP2) and P2X7R in KO and WT primary neurons at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 100 µm. J , Immunostaining for a transfection marker (eGFP) and P2X7R at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 200 µm. * p < 0.05. ** p < 0.005. **** p < 0.0001. ns, non-significant. Data presented as Mean±SEM.

    Techniques Used: In Vitro, Amplification, Expressing, Cell Culture, Western Blot, Immunostaining, Marker, Transfection

    Neurons from P2rx7 −/− mice present less dendritic outgrowth. A , Scheme of the in vitro experiment. B , Immunostaining of a neuronal marker (MAP2) and an astroglial marker (GFAP) and of a transfection marker (eGFP) and a neuronal marker (MAP2) as the control for the transfection in a representative control WT primary hippocampal neuron at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 200 µm. C , Representative hippocampal neurons from WT and P2X7R KO mice transfected with and stained for eGFP. D , Sholl analysis showing the number of intersections versus distance from the soma (radius, μm). E , Using HPLC, nucleotide concentrations in the supernatants of WT and KO primary hippocampal neurons were measured. Quantification of the ( F ) cell body area, ( G ) number of primary dendrites, ( H ) number of endings, and ( I ) mean dendritic length. ** p < 0.005. **** p < 0.0001. ns, non-significant. Data presented as Mean±SEM.
    Figure Legend Snippet: Neurons from P2rx7 −/− mice present less dendritic outgrowth. A , Scheme of the in vitro experiment. B , Immunostaining of a neuronal marker (MAP2) and an astroglial marker (GFAP) and of a transfection marker (eGFP) and a neuronal marker (MAP2) as the control for the transfection in a representative control WT primary hippocampal neuron at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 200 µm. C , Representative hippocampal neurons from WT and P2X7R KO mice transfected with and stained for eGFP. D , Sholl analysis showing the number of intersections versus distance from the soma (radius, μm). E , Using HPLC, nucleotide concentrations in the supernatants of WT and KO primary hippocampal neurons were measured. Quantification of the ( F ) cell body area, ( G ) number of primary dendrites, ( H ) number of endings, and ( I ) mean dendritic length. ** p < 0.005. **** p < 0.0001. ns, non-significant. Data presented as Mean±SEM.

    Techniques Used: In Vitro, Immunostaining, Marker, Transfection, Staining

    Fine-tuning effect of P2X7R on dendritic outgrowth. Antagonists disrupt normal dendritic outgrowth in vitro . A , Scheme of the in vitro experiment. B , Sholl analysis showing the number of intersections versus distance from the soma. Quantification of the ( C ) cell body area, ( D ) number of primary dendrites, ( E ) number of endings, and ( F ) mean dendritic length. Neurons acutely treated with ARL 67156 (100 U/ml) present elongation of dendritic endings. G , Scheme of the in vitro experiment. H , Sholl analysis showing the number of intersections versus distance from the soma. I , Using HPLC, nucleotide concentrations in the supernatants of WT and ARL 67156-treated WT primary hippocampal neurons were measured. Quantification of the ( J ) cell body area, ( K ) number of primary dendrites, ( L ) number of endings, and ( M ) mean total dendritic length. * p < 0.05. ** p < 0.005. *** p < 0.0005. **** p < 0.0001. ns, non-significant. Data presented as Mean±SEM.
    Figure Legend Snippet: Fine-tuning effect of P2X7R on dendritic outgrowth. Antagonists disrupt normal dendritic outgrowth in vitro . A , Scheme of the in vitro experiment. B , Sholl analysis showing the number of intersections versus distance from the soma. Quantification of the ( C ) cell body area, ( D ) number of primary dendrites, ( E ) number of endings, and ( F ) mean dendritic length. Neurons acutely treated with ARL 67156 (100 U/ml) present elongation of dendritic endings. G , Scheme of the in vitro experiment. H , Sholl analysis showing the number of intersections versus distance from the soma. I , Using HPLC, nucleotide concentrations in the supernatants of WT and ARL 67156-treated WT primary hippocampal neurons were measured. Quantification of the ( J ) cell body area, ( K ) number of primary dendrites, ( L ) number of endings, and ( M ) mean total dendritic length. * p < 0.05. ** p < 0.005. *** p < 0.0005. **** p < 0.0001. ns, non-significant. Data presented as Mean±SEM.

    Techniques Used: In Vitro

    Pyramidal neurons from P2rx7 −/− mice present less dendritic outgrowth in the CA1 region of the hippocampus. A , Representative pyramidal hippocampal neurons in slices of the hippocampal CA1 region from P20-29 WT and P2X7R KO mice. B , Sholl analysis showing the number of intersections versus distance from the soma for WT and KO pyramidal neurons. Quantification of the ( C ) cell body area, ( D ) number of primary dendrites, ( E ) number of endings, and ( F ) mean dendritic length. G , Representative pyramidal hippocampal neurons in slices of the hippocampal CA3 region from P20-29 WT and KO mice. H , Sholl analysis showing the number of intersections versus distance from the soma for WT and KO pyramidal neurons. Data are mean ± SEM and analyzed using two-way ANOVA followed by Bonferroni's multiple comparison test. Quantification of the ( I ) cell body area, ( J ) number of primary dendrites, ( K ) number of endings, and ( L ) mean dendritic length. Morphologic deficits correlated with observed deficits in cognitive performance in younger animals. Different aspects of episodic memory, such as ( M ) OL and ( N ) TOR, were analyzed. Deficits in cognitive performance were observed according to both ( O ) the OL index (%) and ( P ) the TOR index (%). Q , Scheme of the experiment. NOR was analyzed ( R ) 2 min after the exploration phase and ( S ) after a longer delay of 24 h. ** p < 0.005. **** p < 0.0001. * p < 0.05. ns, non-significant. Data presented as Mean±SEM.
    Figure Legend Snippet: Pyramidal neurons from P2rx7 −/− mice present less dendritic outgrowth in the CA1 region of the hippocampus. A , Representative pyramidal hippocampal neurons in slices of the hippocampal CA1 region from P20-29 WT and P2X7R KO mice. B , Sholl analysis showing the number of intersections versus distance from the soma for WT and KO pyramidal neurons. Quantification of the ( C ) cell body area, ( D ) number of primary dendrites, ( E ) number of endings, and ( F ) mean dendritic length. G , Representative pyramidal hippocampal neurons in slices of the hippocampal CA3 region from P20-29 WT and KO mice. H , Sholl analysis showing the number of intersections versus distance from the soma for WT and KO pyramidal neurons. Data are mean ± SEM and analyzed using two-way ANOVA followed by Bonferroni's multiple comparison test. Quantification of the ( I ) cell body area, ( J ) number of primary dendrites, ( K ) number of endings, and ( L ) mean dendritic length. Morphologic deficits correlated with observed deficits in cognitive performance in younger animals. Different aspects of episodic memory, such as ( M ) OL and ( N ) TOR, were analyzed. Deficits in cognitive performance were observed according to both ( O ) the OL index (%) and ( P ) the TOR index (%). Q , Scheme of the experiment. NOR was analyzed ( R ) 2 min after the exploration phase and ( S ) after a longer delay of 24 h. ** p < 0.005. **** p < 0.0001. * p < 0.05. ns, non-significant. Data presented as Mean±SEM.

    Techniques Used:

    P2X7R-deficient mice present weaker synaptic strength. A , Representative traces from the WT and KO groups in the presence of gabazine. B , AMPA/NMDA ratio. C , AMPA-mediated current amplitude. D , NMDA-mediated current amplitude. E , AMPA-mediated current rise time. F , AMPA-mediated current decay time. G , NMDA-mediated current rise time. H , NMDA-mediated current decay time. I , Number of spines on WT and KO neurons. J , Representative segments of WT and KO neurons used to count spines. Scale bar, 50 µm. ** p < 0.005. *** p < 0.0005. ns, non-significant. Data presented as Mean±SEM.
    Figure Legend Snippet: P2X7R-deficient mice present weaker synaptic strength. A , Representative traces from the WT and KO groups in the presence of gabazine. B , AMPA/NMDA ratio. C , AMPA-mediated current amplitude. D , NMDA-mediated current amplitude. E , AMPA-mediated current rise time. F , AMPA-mediated current decay time. G , NMDA-mediated current rise time. H , NMDA-mediated current decay time. I , Number of spines on WT and KO neurons. J , Representative segments of WT and KO neurons used to count spines. Scale bar, 50 µm. ** p < 0.005. *** p < 0.0005. ns, non-significant. Data presented as Mean±SEM.

    Techniques Used:

    P2X7R plays a role in a neurodevelopmental model of schizophrenia. A , B , Sholl analysis showing the number of intersections versus distance from the soma for WT neurons, WT neurons treated with different doses of saline or PIC on E12.5, and KO neurons. Quantification of the ( C ) cell body area, ( D ) number of primary dendrites, ( E ) number of endings, and ( F ) mean dendritic length for WT neurons after different treatments. Quantification of the ( G ) cell body area, ( H ) number of primary dendrites, ( I ) number of endings, and ( J ) mean dendritic length for KO neurons after different treatments. K , Using HPLC, nucleotide concentrations in the supernatants of WT and PIC-treated WT primary hippocampal neurons were measured. L , Using HPLC, nucleotide concentrations in the supernatants of KO- and PIC-treated KO primary hippocampal neurons were measured. *** p < 0.0005. **** p < 0.0001. Data presented as Mean±SEM.
    Figure Legend Snippet: P2X7R plays a role in a neurodevelopmental model of schizophrenia. A , B , Sholl analysis showing the number of intersections versus distance from the soma for WT neurons, WT neurons treated with different doses of saline or PIC on E12.5, and KO neurons. Quantification of the ( C ) cell body area, ( D ) number of primary dendrites, ( E ) number of endings, and ( F ) mean dendritic length for WT neurons after different treatments. Quantification of the ( G ) cell body area, ( H ) number of primary dendrites, ( I ) number of endings, and ( J ) mean dendritic length for KO neurons after different treatments. K , Using HPLC, nucleotide concentrations in the supernatants of WT and PIC-treated WT primary hippocampal neurons were measured. L , Using HPLC, nucleotide concentrations in the supernatants of KO- and PIC-treated KO primary hippocampal neurons were measured. *** p < 0.0005. **** p < 0.0001. Data presented as Mean±SEM.

    Techniques Used:

    P2X7R drives PIC-induced schizophrenic-like behavior in mice. A , Overview of the experimental protocol. B , Maternal PIC induced elevation in the level of IL-1β in fetal brain samples of WT mice, while this effect was not observed in samples from KO. Values measured are expressed in picograms per mg of total protein. C , Total distance moved (cm) and ( D ) velocity (cm/s) in the open field were not different between WT animals and PIC-treated WT animals. Quantification of ( E ) the spontaneous alternation percentage and ( F ) NOR percentage in the T-maze test showing that MIA-exposed WT animals exhibited cognitive deficits. G , The amounts of time spent exploring the different cages (seconds) in the social preference test were compared between animals of different genotypes and animal subjected to different treatments. H , The startle response was not different between WT and PIC-treated WT animals. I , PPI was disrupted in PIC-treated WT animals. * p < 0.05. ** p < 0.005. *** p < 0.0005. Data presented as Mean±SEM.
    Figure Legend Snippet: P2X7R drives PIC-induced schizophrenic-like behavior in mice. A , Overview of the experimental protocol. B , Maternal PIC induced elevation in the level of IL-1β in fetal brain samples of WT mice, while this effect was not observed in samples from KO. Values measured are expressed in picograms per mg of total protein. C , Total distance moved (cm) and ( D ) velocity (cm/s) in the open field were not different between WT animals and PIC-treated WT animals. Quantification of ( E ) the spontaneous alternation percentage and ( F ) NOR percentage in the T-maze test showing that MIA-exposed WT animals exhibited cognitive deficits. G , The amounts of time spent exploring the different cages (seconds) in the social preference test were compared between animals of different genotypes and animal subjected to different treatments. H , The startle response was not different between WT and PIC-treated WT animals. I , PPI was disrupted in PIC-treated WT animals. * p < 0.05. ** p < 0.005. *** p < 0.0005. Data presented as Mean±SEM.

    Techniques Used:

    Statistical analysis for molecular and morphologic analysis
    Figure Legend Snippet: Statistical analysis for molecular and morphologic analysis

    Techniques Used: Western Blot, Imaging, Enzyme-linked Immunosorbent Assay

    anti p2x7r  (Alomone Labs)


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    Alomone Labs anti p2x7r
    <t>P2X7R</t> is present in neurons cultivated in vitro during development. A , Scheme of the experiment. B , Scheme of the regions amplified by qPCR. C , Analysis of the expression of extracellular and ( D ) intracellular sequences of P2X7R by real-time qPCR (DIV1) on each DIV. Relative expression is presented as the fold change normalized to GAPDH expression. E , WT cells were cultured for the indicated period of time, and P2X7R protein expression was determined by immunoblotting. Graphs represent densitometric evaluation of the target protein band normalized to the total amount of loaded protein ( n = 4). F , To validate the specificity of the P2X7R antibody, WT and P2X7R-KO cells were cultured for the indicated period of time, and P2X7R protein expression was determined by immunoblotting. Graphs represent the densitometric evaluation of the target protein band normalized to total amount of loaded protein ( n = 4). G , Areas under the curve for WT and KO primary pyramidal cells at DIV4 after acute BzATP (1 m m ) application. H , Areas under the curve for WT and KO primary pyramidal cells at DIV4 after acute calcium ionophore (1 m m ) application. I , Immunostaining of a neuronal marker (MAP2) and P2X7R in KO and WT primary neurons at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 100 µm. J , Immunostaining for a transfection marker (eGFP) and P2X7R at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 200 µm. * p < 0.05. ** p < 0.005. **** p < 0.0001. ns, non-significant. Data presented as Mean±SEM.
    Anti P2x7r, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p2x7r/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti p2x7r - by Bioz Stars, 2023-03
    86/100 stars

    Images

    1) Product Images from "Dual Role of the P2X7 Receptor in Dendritic Outgrowth during Physiological and Pathological Brain Development"

    Article Title: Dual Role of the P2X7 Receptor in Dendritic Outgrowth during Physiological and Pathological Brain Development

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.0805-22.2022

    P2X7R is present in neurons cultivated in vitro during development. A , Scheme of the experiment. B , Scheme of the regions amplified by qPCR. C , Analysis of the expression of extracellular and ( D ) intracellular sequences of P2X7R by real-time qPCR (DIV1) on each DIV. Relative expression is presented as the fold change normalized to GAPDH expression. E , WT cells were cultured for the indicated period of time, and P2X7R protein expression was determined by immunoblotting. Graphs represent densitometric evaluation of the target protein band normalized to the total amount of loaded protein ( n = 4). F , To validate the specificity of the P2X7R antibody, WT and P2X7R-KO cells were cultured for the indicated period of time, and P2X7R protein expression was determined by immunoblotting. Graphs represent the densitometric evaluation of the target protein band normalized to total amount of loaded protein ( n = 4). G , Areas under the curve for WT and KO primary pyramidal cells at DIV4 after acute BzATP (1 m m ) application. H , Areas under the curve for WT and KO primary pyramidal cells at DIV4 after acute calcium ionophore (1 m m ) application. I , Immunostaining of a neuronal marker (MAP2) and P2X7R in KO and WT primary neurons at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 100 µm. J , Immunostaining for a transfection marker (eGFP) and P2X7R at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 200 µm. * p < 0.05. ** p < 0.005. **** p < 0.0001. ns, non-significant. Data presented as Mean±SEM.
    Figure Legend Snippet: P2X7R is present in neurons cultivated in vitro during development. A , Scheme of the experiment. B , Scheme of the regions amplified by qPCR. C , Analysis of the expression of extracellular and ( D ) intracellular sequences of P2X7R by real-time qPCR (DIV1) on each DIV. Relative expression is presented as the fold change normalized to GAPDH expression. E , WT cells were cultured for the indicated period of time, and P2X7R protein expression was determined by immunoblotting. Graphs represent densitometric evaluation of the target protein band normalized to the total amount of loaded protein ( n = 4). F , To validate the specificity of the P2X7R antibody, WT and P2X7R-KO cells were cultured for the indicated period of time, and P2X7R protein expression was determined by immunoblotting. Graphs represent the densitometric evaluation of the target protein band normalized to total amount of loaded protein ( n = 4). G , Areas under the curve for WT and KO primary pyramidal cells at DIV4 after acute BzATP (1 m m ) application. H , Areas under the curve for WT and KO primary pyramidal cells at DIV4 after acute calcium ionophore (1 m m ) application. I , Immunostaining of a neuronal marker (MAP2) and P2X7R in KO and WT primary neurons at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 100 µm. J , Immunostaining for a transfection marker (eGFP) and P2X7R at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 200 µm. * p < 0.05. ** p < 0.005. **** p < 0.0001. ns, non-significant. Data presented as Mean±SEM.

    Techniques Used: In Vitro, Amplification, Expressing, Cell Culture, Western Blot, Immunostaining, Marker, Transfection

    Neurons from P2rx7 −/− mice present less dendritic outgrowth. A , Scheme of the in vitro experiment. B , Immunostaining of a neuronal marker (MAP2) and an astroglial marker (GFAP) and of a transfection marker (eGFP) and a neuronal marker (MAP2) as the control for the transfection in a representative control WT primary hippocampal neuron at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 200 µm. C , Representative hippocampal neurons from WT and P2X7R KO mice transfected with and stained for eGFP. D , Sholl analysis showing the number of intersections versus distance from the soma (radius, μm). E , Using HPLC, nucleotide concentrations in the supernatants of WT and KO primary hippocampal neurons were measured. Quantification of the ( F ) cell body area, ( G ) number of primary dendrites, ( H ) number of endings, and ( I ) mean dendritic length. ** p < 0.005. **** p < 0.0001. ns, non-significant. Data presented as Mean±SEM.
    Figure Legend Snippet: Neurons from P2rx7 −/− mice present less dendritic outgrowth. A , Scheme of the in vitro experiment. B , Immunostaining of a neuronal marker (MAP2) and an astroglial marker (GFAP) and of a transfection marker (eGFP) and a neuronal marker (MAP2) as the control for the transfection in a representative control WT primary hippocampal neuron at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 200 µm. C , Representative hippocampal neurons from WT and P2X7R KO mice transfected with and stained for eGFP. D , Sholl analysis showing the number of intersections versus distance from the soma (radius, μm). E , Using HPLC, nucleotide concentrations in the supernatants of WT and KO primary hippocampal neurons were measured. Quantification of the ( F ) cell body area, ( G ) number of primary dendrites, ( H ) number of endings, and ( I ) mean dendritic length. ** p < 0.005. **** p < 0.0001. ns, non-significant. Data presented as Mean±SEM.

    Techniques Used: In Vitro, Immunostaining, Marker, Transfection, Staining

    Fine-tuning effect of P2X7R on dendritic outgrowth. Antagonists disrupt normal dendritic outgrowth in vitro . A , Scheme of the in vitro experiment. B , Sholl analysis showing the number of intersections versus distance from the soma. Quantification of the ( C ) cell body area, ( D ) number of primary dendrites, ( E ) number of endings, and ( F ) mean dendritic length. Neurons acutely treated with ARL 67156 (100 U/ml) present elongation of dendritic endings. G , Scheme of the in vitro experiment. H , Sholl analysis showing the number of intersections versus distance from the soma. I , Using HPLC, nucleotide concentrations in the supernatants of WT and ARL 67156-treated WT primary hippocampal neurons were measured. Quantification of the ( J ) cell body area, ( K ) number of primary dendrites, ( L ) number of endings, and ( M ) mean total dendritic length. * p < 0.05. ** p < 0.005. *** p < 0.0005. **** p < 0.0001. ns, non-significant. Data presented as Mean±SEM.
    Figure Legend Snippet: Fine-tuning effect of P2X7R on dendritic outgrowth. Antagonists disrupt normal dendritic outgrowth in vitro . A , Scheme of the in vitro experiment. B , Sholl analysis showing the number of intersections versus distance from the soma. Quantification of the ( C ) cell body area, ( D ) number of primary dendrites, ( E ) number of endings, and ( F ) mean dendritic length. Neurons acutely treated with ARL 67156 (100 U/ml) present elongation of dendritic endings. G , Scheme of the in vitro experiment. H , Sholl analysis showing the number of intersections versus distance from the soma. I , Using HPLC, nucleotide concentrations in the supernatants of WT and ARL 67156-treated WT primary hippocampal neurons were measured. Quantification of the ( J ) cell body area, ( K ) number of primary dendrites, ( L ) number of endings, and ( M ) mean total dendritic length. * p < 0.05. ** p < 0.005. *** p < 0.0005. **** p < 0.0001. ns, non-significant. Data presented as Mean±SEM.

    Techniques Used: In Vitro

    Pyramidal neurons from P2rx7 −/− mice present less dendritic outgrowth in the CA1 region of the hippocampus. A , Representative pyramidal hippocampal neurons in slices of the hippocampal CA1 region from P20-29 WT and P2X7R KO mice. B , Sholl analysis showing the number of intersections versus distance from the soma for WT and KO pyramidal neurons. Quantification of the ( C ) cell body area, ( D ) number of primary dendrites, ( E ) number of endings, and ( F ) mean dendritic length. G , Representative pyramidal hippocampal neurons in slices of the hippocampal CA3 region from P20-29 WT and KO mice. H , Sholl analysis showing the number of intersections versus distance from the soma for WT and KO pyramidal neurons. Data are mean ± SEM and analyzed using two-way ANOVA followed by Bonferroni's multiple comparison test. Quantification of the ( I ) cell body area, ( J ) number of primary dendrites, ( K ) number of endings, and ( L ) mean dendritic length. Morphologic deficits correlated with observed deficits in cognitive performance in younger animals. Different aspects of episodic memory, such as ( M ) OL and ( N ) TOR, were analyzed. Deficits in cognitive performance were observed according to both ( O ) the OL index (%) and ( P ) the TOR index (%). Q , Scheme of the experiment. NOR was analyzed ( R ) 2 min after the exploration phase and ( S ) after a longer delay of 24 h. ** p < 0.005. **** p < 0.0001. * p < 0.05. ns, non-significant. Data presented as Mean±SEM.
    Figure Legend Snippet: Pyramidal neurons from P2rx7 −/− mice present less dendritic outgrowth in the CA1 region of the hippocampus. A , Representative pyramidal hippocampal neurons in slices of the hippocampal CA1 region from P20-29 WT and P2X7R KO mice. B , Sholl analysis showing the number of intersections versus distance from the soma for WT and KO pyramidal neurons. Quantification of the ( C ) cell body area, ( D ) number of primary dendrites, ( E ) number of endings, and ( F ) mean dendritic length. G , Representative pyramidal hippocampal neurons in slices of the hippocampal CA3 region from P20-29 WT and KO mice. H , Sholl analysis showing the number of intersections versus distance from the soma for WT and KO pyramidal neurons. Data are mean ± SEM and analyzed using two-way ANOVA followed by Bonferroni's multiple comparison test. Quantification of the ( I ) cell body area, ( J ) number of primary dendrites, ( K ) number of endings, and ( L ) mean dendritic length. Morphologic deficits correlated with observed deficits in cognitive performance in younger animals. Different aspects of episodic memory, such as ( M ) OL and ( N ) TOR, were analyzed. Deficits in cognitive performance were observed according to both ( O ) the OL index (%) and ( P ) the TOR index (%). Q , Scheme of the experiment. NOR was analyzed ( R ) 2 min after the exploration phase and ( S ) after a longer delay of 24 h. ** p < 0.005. **** p < 0.0001. * p < 0.05. ns, non-significant. Data presented as Mean±SEM.

    Techniques Used:

    P2X7R-deficient mice present weaker synaptic strength. A , Representative traces from the WT and KO groups in the presence of gabazine. B , AMPA/NMDA ratio. C , AMPA-mediated current amplitude. D , NMDA-mediated current amplitude. E , AMPA-mediated current rise time. F , AMPA-mediated current decay time. G , NMDA-mediated current rise time. H , NMDA-mediated current decay time. I , Number of spines on WT and KO neurons. J , Representative segments of WT and KO neurons used to count spines. Scale bar, 50 µm. ** p < 0.005. *** p < 0.0005. ns, non-significant. Data presented as Mean±SEM.
    Figure Legend Snippet: P2X7R-deficient mice present weaker synaptic strength. A , Representative traces from the WT and KO groups in the presence of gabazine. B , AMPA/NMDA ratio. C , AMPA-mediated current amplitude. D , NMDA-mediated current amplitude. E , AMPA-mediated current rise time. F , AMPA-mediated current decay time. G , NMDA-mediated current rise time. H , NMDA-mediated current decay time. I , Number of spines on WT and KO neurons. J , Representative segments of WT and KO neurons used to count spines. Scale bar, 50 µm. ** p < 0.005. *** p < 0.0005. ns, non-significant. Data presented as Mean±SEM.

    Techniques Used:

    P2X7R plays a role in a neurodevelopmental model of schizophrenia. A , B , Sholl analysis showing the number of intersections versus distance from the soma for WT neurons, WT neurons treated with different doses of saline or PIC on E12.5, and KO neurons. Quantification of the ( C ) cell body area, ( D ) number of primary dendrites, ( E ) number of endings, and ( F ) mean dendritic length for WT neurons after different treatments. Quantification of the ( G ) cell body area, ( H ) number of primary dendrites, ( I ) number of endings, and ( J ) mean dendritic length for KO neurons after different treatments. K , Using HPLC, nucleotide concentrations in the supernatants of WT and PIC-treated WT primary hippocampal neurons were measured. L , Using HPLC, nucleotide concentrations in the supernatants of KO- and PIC-treated KO primary hippocampal neurons were measured. *** p < 0.0005. **** p < 0.0001. Data presented as Mean±SEM.
    Figure Legend Snippet: P2X7R plays a role in a neurodevelopmental model of schizophrenia. A , B , Sholl analysis showing the number of intersections versus distance from the soma for WT neurons, WT neurons treated with different doses of saline or PIC on E12.5, and KO neurons. Quantification of the ( C ) cell body area, ( D ) number of primary dendrites, ( E ) number of endings, and ( F ) mean dendritic length for WT neurons after different treatments. Quantification of the ( G ) cell body area, ( H ) number of primary dendrites, ( I ) number of endings, and ( J ) mean dendritic length for KO neurons after different treatments. K , Using HPLC, nucleotide concentrations in the supernatants of WT and PIC-treated WT primary hippocampal neurons were measured. L , Using HPLC, nucleotide concentrations in the supernatants of KO- and PIC-treated KO primary hippocampal neurons were measured. *** p < 0.0005. **** p < 0.0001. Data presented as Mean±SEM.

    Techniques Used:

    P2X7R drives PIC-induced schizophrenic-like behavior in mice. A , Overview of the experimental protocol. B , Maternal PIC induced elevation in the level of IL-1β in fetal brain samples of WT mice, while this effect was not observed in samples from KO. Values measured are expressed in picograms per mg of total protein. C , Total distance moved (cm) and ( D ) velocity (cm/s) in the open field were not different between WT animals and PIC-treated WT animals. Quantification of ( E ) the spontaneous alternation percentage and ( F ) NOR percentage in the T-maze test showing that MIA-exposed WT animals exhibited cognitive deficits. G , The amounts of time spent exploring the different cages (seconds) in the social preference test were compared between animals of different genotypes and animal subjected to different treatments. H , The startle response was not different between WT and PIC-treated WT animals. I , PPI was disrupted in PIC-treated WT animals. * p < 0.05. ** p < 0.005. *** p < 0.0005. Data presented as Mean±SEM.
    Figure Legend Snippet: P2X7R drives PIC-induced schizophrenic-like behavior in mice. A , Overview of the experimental protocol. B , Maternal PIC induced elevation in the level of IL-1β in fetal brain samples of WT mice, while this effect was not observed in samples from KO. Values measured are expressed in picograms per mg of total protein. C , Total distance moved (cm) and ( D ) velocity (cm/s) in the open field were not different between WT animals and PIC-treated WT animals. Quantification of ( E ) the spontaneous alternation percentage and ( F ) NOR percentage in the T-maze test showing that MIA-exposed WT animals exhibited cognitive deficits. G , The amounts of time spent exploring the different cages (seconds) in the social preference test were compared between animals of different genotypes and animal subjected to different treatments. H , The startle response was not different between WT and PIC-treated WT animals. I , PPI was disrupted in PIC-treated WT animals. * p < 0.05. ** p < 0.005. *** p < 0.0005. Data presented as Mean±SEM.

    Techniques Used:

    Statistical analysis for molecular and morphologic analysis
    Figure Legend Snippet: Statistical analysis for molecular and morphologic analysis

    Techniques Used: Western Blot, Imaging, Enzyme-linked Immunosorbent Assay

    anti p2x7r  (Alomone Labs)


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    Alomone Labs anti p2x7r
    Primary antibodies used for Western Blot (WB) and immunohistochemistry (IHC).
    Anti P2x7r, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p2x7r/product/Alomone Labs
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    96/100 stars

    Images

    1) Product Images from "A Co-Culture System with an Organotypic Lung Slice and an Immortal Alveolar Macrophage Cell Line to Quantify Silica-Induced Inflammation"

    Article Title: A Co-Culture System with an Organotypic Lung Slice and an Immortal Alveolar Macrophage Cell Line to Quantify Silica-Induced Inflammation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0117056

    Primary antibodies used for Western Blot (WB) and immunohistochemistry (IHC).
    Figure Legend Snippet: Primary antibodies used for Western Blot (WB) and immunohistochemistry (IHC).

    Techniques Used: Western Blot, Immunohistochemistry

    p2x7r  (Alomone Labs)


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    Alomone Labs p2x7r
    Sequences of siRNA to <t> P2X7R. </t>
    P2x7r, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p2x7r/product/Alomone Labs
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p2x7r - by Bioz Stars, 2023-03
    96/100 stars

    Images

    1) Product Images from "Inhibition of P2X7 Purinergic Receptor Ameliorates Cardiac Fibrosis by Suppressing NLRP3/IL-1 β Pathway"

    Article Title: Inhibition of P2X7 Purinergic Receptor Ameliorates Cardiac Fibrosis by Suppressing NLRP3/IL-1 β Pathway

    Journal: Oxidative Medicine and Cellular Longevity

    doi: 10.1155/2020/7956274

    Sequences of siRNA to P2X7R.
    Figure Legend Snippet: Sequences of siRNA to P2X7R.

    Techniques Used:

    Rat RT-qPCR primer sequences.
    Figure Legend Snippet: Rat RT-qPCR primer sequences.

    Techniques Used: Sequencing

    Mice RT-qPCR primer sequences.
    Figure Legend Snippet: Mice RT-qPCR primer sequences.

    Techniques Used: Sequencing

    P2X7R was upregulated in the mice heart after TAC surgery. (a) Cardiac images and HW/BW ( n = 8). (b) HE-, Sirius Red-, and Masson's trichrome-stained sections of the hearts following four weeks of TAC treatment ( n = 6). (c) Representative images of an echocardiographic assessment of mice after TAC (4 weeks). Cardiac function indicators measured by echocardiography (LVEF (%), FS (%), LVPWd, IVSd, and LVIDd) in TAC- and sham-operated mice. (d) Expression of CTGF, periostin, and α -SMA mRNAs and proteins in heart cells ( n = 6). (e) Gene expression of P2X receptors in TAC- and sham-operated mice ( n = 6) at 4 weeks and gene expression of P2X7R in heart cells at 3 days, 1 week, 2 weeks, and 4 weeks after TAC- or sham-operated treatment ( n = 6). (f) Expression of P2X7R protein in mouse heart cells ( n = 6). Results are presented as means ± standard error of the mean. ∗ indicates P < 0.05, ∗∗ indicates P < 0.01, and ∗∗∗ indicates P < 0.001 versus sham.
    Figure Legend Snippet: P2X7R was upregulated in the mice heart after TAC surgery. (a) Cardiac images and HW/BW ( n = 8). (b) HE-, Sirius Red-, and Masson's trichrome-stained sections of the hearts following four weeks of TAC treatment ( n = 6). (c) Representative images of an echocardiographic assessment of mice after TAC (4 weeks). Cardiac function indicators measured by echocardiography (LVEF (%), FS (%), LVPWd, IVSd, and LVIDd) in TAC- and sham-operated mice. (d) Expression of CTGF, periostin, and α -SMA mRNAs and proteins in heart cells ( n = 6). (e) Gene expression of P2X receptors in TAC- and sham-operated mice ( n = 6) at 4 weeks and gene expression of P2X7R in heart cells at 3 days, 1 week, 2 weeks, and 4 weeks after TAC- or sham-operated treatment ( n = 6). (f) Expression of P2X7R protein in mouse heart cells ( n = 6). Results are presented as means ± standard error of the mean. ∗ indicates P < 0.05, ∗∗ indicates P < 0.01, and ∗∗∗ indicates P < 0.001 versus sham.

    Techniques Used: Staining, Expressing

    TGF- β 1-treatment upregulated P2X7R in CFs. (a) Immunofluorescence images showing α -SMA expression in CFs ( α -SMA (green) and nuclei DAPI (blue), n = 300). (b) Rate of cell proliferation (EdU-positive cells (red) and nuclei DAPI (blue), n = 120). (c) Expression of TGF- β , α -SMA, CTGF, and periostin mRNAs ( n = 3). (d) Expression of CTGF, α -SMA, periostin, and TGF- β proteins ( n = 3). (e) Western blot results showing P2X7R expression in CFs ( n = 3). (f) Immunofluorescence images showing P2RX7 expression in CFs (P2X7R (red) and nuclei DAPI (blue), n = 60). Results are presented as means ± standard error of the mean; ∗ indicates P < 0.05, ∗∗ indicates P < 0.01, and ∗∗∗ indicates P < 0.001 versus control.
    Figure Legend Snippet: TGF- β 1-treatment upregulated P2X7R in CFs. (a) Immunofluorescence images showing α -SMA expression in CFs ( α -SMA (green) and nuclei DAPI (blue), n = 300). (b) Rate of cell proliferation (EdU-positive cells (red) and nuclei DAPI (blue), n = 120). (c) Expression of TGF- β , α -SMA, CTGF, and periostin mRNAs ( n = 3). (d) Expression of CTGF, α -SMA, periostin, and TGF- β proteins ( n = 3). (e) Western blot results showing P2X7R expression in CFs ( n = 3). (f) Immunofluorescence images showing P2RX7 expression in CFs (P2X7R (red) and nuclei DAPI (blue), n = 60). Results are presented as means ± standard error of the mean; ∗ indicates P < 0.05, ∗∗ indicates P < 0.01, and ∗∗∗ indicates P < 0.001 versus control.

    Techniques Used: Immunofluorescence, Expressing, Western Blot

    Knockdown of P2X7R alleviates the effects of TGF- β 1 on CF activation. (a) Expression of P2X7R mRNA after siP2X7R transfection ( n = 6). (b) Expression of collagen I, CTGF, α -SMA, and TGF- β mRNAs after transfection of CFs with siP2X7R ( n = 6). (c, d) Expression of periostin, α -SMA, CTGF, P2X7R, NLRP3, IL-1 β , and caspase-1 proteins after transfection of CFs with siP2X7R ( n = 6). (e) Expression of α -SMA in CFs by immunofluorescence staining ( α -SMA (green) and nuclei DAPI (blue), n = 300). (f) Changes in CF proliferation (EdU-positive cells (red) and nuclei DAPI (blue), n = 120). (g) Expression of collagen I, CTGF, α -SMA, and TGF- β mRNAs after treatment of CFs with BBG ( n = 6). (h) Expression of P2X7R protein after BBG treatment ( n = 6). (i) Expression of periostin, α -SMA, CTGF, NLRP3, IL-1 β , and caspase-1 proteins after BBG treatment ( n = 6). (j) Expression of periostin, α -SMA, TGF- β and CTGF proteins in activated CFs after BzATP treatment ( n = 6). Results are presented as means ± standard error of the mean; ∗ indicates P < 0.05, ∗∗ indicates P < 0.01, and ∗∗∗ indicates P < 0.001 versus control or siNC; # indicates P < 0.05, ## indicates P < 0.01, and ### indicates P < 0.001.
    Figure Legend Snippet: Knockdown of P2X7R alleviates the effects of TGF- β 1 on CF activation. (a) Expression of P2X7R mRNA after siP2X7R transfection ( n = 6). (b) Expression of collagen I, CTGF, α -SMA, and TGF- β mRNAs after transfection of CFs with siP2X7R ( n = 6). (c, d) Expression of periostin, α -SMA, CTGF, P2X7R, NLRP3, IL-1 β , and caspase-1 proteins after transfection of CFs with siP2X7R ( n = 6). (e) Expression of α -SMA in CFs by immunofluorescence staining ( α -SMA (green) and nuclei DAPI (blue), n = 300). (f) Changes in CF proliferation (EdU-positive cells (red) and nuclei DAPI (blue), n = 120). (g) Expression of collagen I, CTGF, α -SMA, and TGF- β mRNAs after treatment of CFs with BBG ( n = 6). (h) Expression of P2X7R protein after BBG treatment ( n = 6). (i) Expression of periostin, α -SMA, CTGF, NLRP3, IL-1 β , and caspase-1 proteins after BBG treatment ( n = 6). (j) Expression of periostin, α -SMA, TGF- β and CTGF proteins in activated CFs after BzATP treatment ( n = 6). Results are presented as means ± standard error of the mean; ∗ indicates P < 0.05, ∗∗ indicates P < 0.01, and ∗∗∗ indicates P < 0.001 versus control or siNC; # indicates P < 0.05, ## indicates P < 0.01, and ### indicates P < 0.001.

    Techniques Used: Activation Assay, Expressing, Transfection, Immunofluorescence, Staining

    A schematic illustration of the mechanism of P2X7R in pressure overload-induced cardiac fibrosis and TGF- β 1-induced CF activation. In TAC-induced injury, pathological stress, such as pressure overload and TGF- β 1, triggers the release of ATP. Elevated ATP activates P2X7 receptors, contributing to P2X7-mediated NLRP3 inflammasome (NLRP3, ASC, and caspase-1) activation. This induces the release of IL-1 β , causing severe inflammation that precipitates cardiac fibrosis and aggravates TGF- β 1-induced CF activation. Moreover, inhibition of P2X7R with BBG inhibitor in TAC mice or CFs primed with TGF- β 1 reduces fibrotic extracellular matrix (ECM) gene expression and cardiac fibrosis.
    Figure Legend Snippet: A schematic illustration of the mechanism of P2X7R in pressure overload-induced cardiac fibrosis and TGF- β 1-induced CF activation. In TAC-induced injury, pathological stress, such as pressure overload and TGF- β 1, triggers the release of ATP. Elevated ATP activates P2X7 receptors, contributing to P2X7-mediated NLRP3 inflammasome (NLRP3, ASC, and caspase-1) activation. This induces the release of IL-1 β , causing severe inflammation that precipitates cardiac fibrosis and aggravates TGF- β 1-induced CF activation. Moreover, inhibition of P2X7R with BBG inhibitor in TAC mice or CFs primed with TGF- β 1 reduces fibrotic extracellular matrix (ECM) gene expression and cardiac fibrosis.

    Techniques Used: Activation Assay, Inhibition, Expressing

    p2x7r  (Alomone Labs)


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    Alomone Labs p2x7r
    Spleen cells from 3 to 4-mo-old MRL +/+ ( solid line ), MRL/ lpr ( dashed line ) and P2X7-deficient B6 <t>P2X7R−/−</t> ( dotted line ) mice (n = 3 mice/group) were treated for 45 min at 37°C with doses of ATP ranging from 100 to 5000 µM. Spleen cells were then triple-stained with anti-CD90, anti-CD19 and anti-CD62L mAb to assess by flow cytometry: (A) the percentage of CD62L-expressing CD19 – CD90 + T cells and (B) MFI of CD62L on CD19 – CD90 + T cells. Results are expressed as the mean percentage of initial expression ± SE.
    P2x7r, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p2x7r/product/Alomone Labs
    Average 96 stars, based on 1 article reviews
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    p2x7r - by Bioz Stars, 2023-03
    96/100 stars

    Images

    1) Product Images from "Loss of P2X7 Receptor Plasma Membrane Expression and Function in Pathogenic B220 + Double-Negative T Lymphocytes of Autoimmune MRL/ lpr Mice"

    Article Title: Loss of P2X7 Receptor Plasma Membrane Expression and Function in Pathogenic B220 + Double-Negative T Lymphocytes of Autoimmune MRL/ lpr Mice

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0052161

    Spleen cells from 3 to 4-mo-old MRL +/+ ( solid line ), MRL/ lpr ( dashed line ) and P2X7-deficient B6 P2X7R−/− ( dotted line ) mice (n = 3 mice/group) were treated for 45 min at 37°C with doses of ATP ranging from 100 to 5000 µM. Spleen cells were then triple-stained with anti-CD90, anti-CD19 and anti-CD62L mAb to assess by flow cytometry: (A) the percentage of CD62L-expressing CD19 – CD90 + T cells and (B) MFI of CD62L on CD19 – CD90 + T cells. Results are expressed as the mean percentage of initial expression ± SE.
    Figure Legend Snippet: Spleen cells from 3 to 4-mo-old MRL +/+ ( solid line ), MRL/ lpr ( dashed line ) and P2X7-deficient B6 P2X7R−/− ( dotted line ) mice (n = 3 mice/group) were treated for 45 min at 37°C with doses of ATP ranging from 100 to 5000 µM. Spleen cells were then triple-stained with anti-CD90, anti-CD19 and anti-CD62L mAb to assess by flow cytometry: (A) the percentage of CD62L-expressing CD19 – CD90 + T cells and (B) MFI of CD62L on CD19 – CD90 + T cells. Results are expressed as the mean percentage of initial expression ± SE.

    Techniques Used: Staining, Flow Cytometry, Expressing

    (A) The levels of CD39 expression on B220 – (either CD4 + or CD8 + ) (□) and B220 + DN (▪) T-cell subsets as well as on CD19 + B cells (▪) from MRL/ lpr mice (n = 3) were analyzed by flow cytometry. Results shown on histogram are normalized MFI (nMFI) calculated as MFI of positive cells × percentage of positive cells. Asterisks denote statistically significant differences between B220 + DN T cells or B220 – CD8 + T cells and B220 – CD4 + T cells: * p ≤0.05; ** p ≤0.01. (B) P2X7R mRNA expression was analyzed in FACS-purified B220 – (□) and B220 + (▪) CD19 – CD90 + T-cell subsets from individual B6, MRL +/+ and MRL/ lpr mice by real-time RT-PCR. Specific P2X7R cDNA was quantified by standard curves based on known amounts of PCR-amplified P2X7R cDNA. Data are expressed as amount (pg) of P2X7R RNA per cell, and histograms represent the mean ± SE of three independent experiments (n = 8 mice/group). (C) P2X7R protein levels in whole LN cells from 3 to 4-mo-old MRL +/+ , MRL/ lpr and B6 P2X7−/− mice, and FACS-sorted B220 – and B220 + CD19 – CD90 + T-cell subsets from MRL/ lpr mice were analyzed by western blotting using affinity-purified rabbit anti-P2X7R polyclonal antibodies (1∶1000; Alomone Laboratories). A non-specific band (*) is frequently observed with this polyclonal antibody . The blot was stripped and reprobed with anti-actin mAb. (D and E) Flow cytometric analysis of P2X7R expression levels on B220 – and B220 + T-cell subpopulations. Spleen cells from MRL +/+ , MRL/ lpr and B6 P2X7R−/− mice (n = 3 mice/group) were stained with anti-CD90 mAb, anti-B220 mAb, rabbit polyclonal anti-P2X7R antiserum (1∶100) generated by genetic immunization and with either anti-CD4 mAb, anti-CD8 mAb or anti-CD4 plus anti-CD8 mAbs. (D) Overlay histograms showing the expression levels of P2X7R on gated CD4 T cells or CD8 T cells from MRL +/+ mice (open histogram) and B6 P2X7R−/− mice (shaded histogram). (E) Overlay histograms comparing the expression levels of P2X7R on B220 – (–) and B220 + (–) T cells, either CD4 + ( left ) or CD8 + ( middle ), from MRL/ lpr mice. Right , overlay histogram comparing the expression levels of P2X7R on B220 + DN T cells (–) and B220 – CD4 + T cells (–) from MRL/ lpr mice. The results shown are representative of at least four independent experiments.
    Figure Legend Snippet: (A) The levels of CD39 expression on B220 – (either CD4 + or CD8 + ) (□) and B220 + DN (▪) T-cell subsets as well as on CD19 + B cells (▪) from MRL/ lpr mice (n = 3) were analyzed by flow cytometry. Results shown on histogram are normalized MFI (nMFI) calculated as MFI of positive cells × percentage of positive cells. Asterisks denote statistically significant differences between B220 + DN T cells or B220 – CD8 + T cells and B220 – CD4 + T cells: * p ≤0.05; ** p ≤0.01. (B) P2X7R mRNA expression was analyzed in FACS-purified B220 – (□) and B220 + (▪) CD19 – CD90 + T-cell subsets from individual B6, MRL +/+ and MRL/ lpr mice by real-time RT-PCR. Specific P2X7R cDNA was quantified by standard curves based on known amounts of PCR-amplified P2X7R cDNA. Data are expressed as amount (pg) of P2X7R RNA per cell, and histograms represent the mean ± SE of three independent experiments (n = 8 mice/group). (C) P2X7R protein levels in whole LN cells from 3 to 4-mo-old MRL +/+ , MRL/ lpr and B6 P2X7−/− mice, and FACS-sorted B220 – and B220 + CD19 – CD90 + T-cell subsets from MRL/ lpr mice were analyzed by western blotting using affinity-purified rabbit anti-P2X7R polyclonal antibodies (1∶1000; Alomone Laboratories). A non-specific band (*) is frequently observed with this polyclonal antibody . The blot was stripped and reprobed with anti-actin mAb. (D and E) Flow cytometric analysis of P2X7R expression levels on B220 – and B220 + T-cell subpopulations. Spleen cells from MRL +/+ , MRL/ lpr and B6 P2X7R−/− mice (n = 3 mice/group) were stained with anti-CD90 mAb, anti-B220 mAb, rabbit polyclonal anti-P2X7R antiserum (1∶100) generated by genetic immunization and with either anti-CD4 mAb, anti-CD8 mAb or anti-CD4 plus anti-CD8 mAbs. (D) Overlay histograms showing the expression levels of P2X7R on gated CD4 T cells or CD8 T cells from MRL +/+ mice (open histogram) and B6 P2X7R−/− mice (shaded histogram). (E) Overlay histograms comparing the expression levels of P2X7R on B220 – (–) and B220 + (–) T cells, either CD4 + ( left ) or CD8 + ( middle ), from MRL/ lpr mice. Right , overlay histogram comparing the expression levels of P2X7R on B220 + DN T cells (–) and B220 – CD4 + T cells (–) from MRL/ lpr mice. The results shown are representative of at least four independent experiments.

    Techniques Used: Expressing, Flow Cytometry, Purification, Quantitative RT-PCR, Amplification, Western Blot, Affinity Purification, Staining, Generated

    affinity purified rabbit anti p2x7r antibodies  (Alomone Labs)


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    Alomone Labs affinity purified rabbit anti p2x7r antibodies
    Spleen cells from 3 to 4-mo-old MRL +/+ ( solid line ), MRL/ lpr ( dashed line ) and P2X7-deficient B6 <t>P2X7R−/−</t> ( dotted line ) mice (n = 3 mice/group) were treated for 45 min at 37°C with doses of ATP ranging from 100 to 5000 µM. Spleen cells were then triple-stained with anti-CD90, anti-CD19 and anti-CD62L mAb to assess by flow cytometry: (A) the percentage of CD62L-expressing CD19 – CD90 + T cells and (B) MFI of CD62L on CD19 – CD90 + T cells. Results are expressed as the mean percentage of initial expression ± SE.
    Affinity Purified Rabbit Anti P2x7r Antibodies, 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
    https://www.bioz.com/result/affinity purified rabbit anti p2x7r antibodies/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    affinity purified rabbit anti p2x7r antibodies - by Bioz Stars, 2023-03
    93/100 stars

    Images

    1) Product Images from "Loss of P2X7 Receptor Plasma Membrane Expression and Function in Pathogenic B220 + Double-Negative T Lymphocytes of Autoimmune MRL/ lpr Mice"

    Article Title: Loss of P2X7 Receptor Plasma Membrane Expression and Function in Pathogenic B220 + Double-Negative T Lymphocytes of Autoimmune MRL/ lpr Mice

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0052161

    Spleen cells from 3 to 4-mo-old MRL +/+ ( solid line ), MRL/ lpr ( dashed line ) and P2X7-deficient B6 P2X7R−/− ( dotted line ) mice (n = 3 mice/group) were treated for 45 min at 37°C with doses of ATP ranging from 100 to 5000 µM. Spleen cells were then triple-stained with anti-CD90, anti-CD19 and anti-CD62L mAb to assess by flow cytometry: (A) the percentage of CD62L-expressing CD19 – CD90 + T cells and (B) MFI of CD62L on CD19 – CD90 + T cells. Results are expressed as the mean percentage of initial expression ± SE.
    Figure Legend Snippet: Spleen cells from 3 to 4-mo-old MRL +/+ ( solid line ), MRL/ lpr ( dashed line ) and P2X7-deficient B6 P2X7R−/− ( dotted line ) mice (n = 3 mice/group) were treated for 45 min at 37°C with doses of ATP ranging from 100 to 5000 µM. Spleen cells were then triple-stained with anti-CD90, anti-CD19 and anti-CD62L mAb to assess by flow cytometry: (A) the percentage of CD62L-expressing CD19 – CD90 + T cells and (B) MFI of CD62L on CD19 – CD90 + T cells. Results are expressed as the mean percentage of initial expression ± SE.

    Techniques Used: Staining, Flow Cytometry, Expressing

    (A) The levels of CD39 expression on B220 – (either CD4 + or CD8 + ) (□) and B220 + DN (▪) T-cell subsets as well as on CD19 + B cells (▪) from MRL/ lpr mice (n = 3) were analyzed by flow cytometry. Results shown on histogram are normalized MFI (nMFI) calculated as MFI of positive cells × percentage of positive cells. Asterisks denote statistically significant differences between B220 + DN T cells or B220 – CD8 + T cells and B220 – CD4 + T cells: * p ≤0.05; ** p ≤0.01. (B) P2X7R mRNA expression was analyzed in FACS-purified B220 – (□) and B220 + (▪) CD19 – CD90 + T-cell subsets from individual B6, MRL +/+ and MRL/ lpr mice by real-time RT-PCR. Specific P2X7R cDNA was quantified by standard curves based on known amounts of PCR-amplified P2X7R cDNA. Data are expressed as amount (pg) of P2X7R RNA per cell, and histograms represent the mean ± SE of three independent experiments (n = 8 mice/group). (C) P2X7R protein levels in whole LN cells from 3 to 4-mo-old MRL +/+ , MRL/ lpr and B6 P2X7−/− mice, and FACS-sorted B220 – and B220 + CD19 – CD90 + T-cell subsets from MRL/ lpr mice were analyzed by western blotting using affinity-purified rabbit anti-P2X7R polyclonal antibodies (1∶1000; Alomone Laboratories). A non-specific band (*) is frequently observed with this polyclonal antibody . The blot was stripped and reprobed with anti-actin mAb. (D and E) Flow cytometric analysis of P2X7R expression levels on B220 – and B220 + T-cell subpopulations. Spleen cells from MRL +/+ , MRL/ lpr and B6 P2X7R−/− mice (n = 3 mice/group) were stained with anti-CD90 mAb, anti-B220 mAb, rabbit polyclonal anti-P2X7R antiserum (1∶100) generated by genetic immunization and with either anti-CD4 mAb, anti-CD8 mAb or anti-CD4 plus anti-CD8 mAbs. (D) Overlay histograms showing the expression levels of P2X7R on gated CD4 T cells or CD8 T cells from MRL +/+ mice (open histogram) and B6 P2X7R−/− mice (shaded histogram). (E) Overlay histograms comparing the expression levels of P2X7R on B220 – (–) and B220 + (–) T cells, either CD4 + ( left ) or CD8 + ( middle ), from MRL/ lpr mice. Right , overlay histogram comparing the expression levels of P2X7R on B220 + DN T cells (–) and B220 – CD4 + T cells (–) from MRL/ lpr mice. The results shown are representative of at least four independent experiments.
    Figure Legend Snippet: (A) The levels of CD39 expression on B220 – (either CD4 + or CD8 + ) (□) and B220 + DN (▪) T-cell subsets as well as on CD19 + B cells (▪) from MRL/ lpr mice (n = 3) were analyzed by flow cytometry. Results shown on histogram are normalized MFI (nMFI) calculated as MFI of positive cells × percentage of positive cells. Asterisks denote statistically significant differences between B220 + DN T cells or B220 – CD8 + T cells and B220 – CD4 + T cells: * p ≤0.05; ** p ≤0.01. (B) P2X7R mRNA expression was analyzed in FACS-purified B220 – (□) and B220 + (▪) CD19 – CD90 + T-cell subsets from individual B6, MRL +/+ and MRL/ lpr mice by real-time RT-PCR. Specific P2X7R cDNA was quantified by standard curves based on known amounts of PCR-amplified P2X7R cDNA. Data are expressed as amount (pg) of P2X7R RNA per cell, and histograms represent the mean ± SE of three independent experiments (n = 8 mice/group). (C) P2X7R protein levels in whole LN cells from 3 to 4-mo-old MRL +/+ , MRL/ lpr and B6 P2X7−/− mice, and FACS-sorted B220 – and B220 + CD19 – CD90 + T-cell subsets from MRL/ lpr mice were analyzed by western blotting using affinity-purified rabbit anti-P2X7R polyclonal antibodies (1∶1000; Alomone Laboratories). A non-specific band (*) is frequently observed with this polyclonal antibody . The blot was stripped and reprobed with anti-actin mAb. (D and E) Flow cytometric analysis of P2X7R expression levels on B220 – and B220 + T-cell subpopulations. Spleen cells from MRL +/+ , MRL/ lpr and B6 P2X7R−/− mice (n = 3 mice/group) were stained with anti-CD90 mAb, anti-B220 mAb, rabbit polyclonal anti-P2X7R antiserum (1∶100) generated by genetic immunization and with either anti-CD4 mAb, anti-CD8 mAb or anti-CD4 plus anti-CD8 mAbs. (D) Overlay histograms showing the expression levels of P2X7R on gated CD4 T cells or CD8 T cells from MRL +/+ mice (open histogram) and B6 P2X7R−/− mice (shaded histogram). (E) Overlay histograms comparing the expression levels of P2X7R on B220 – (–) and B220 + (–) T cells, either CD4 + ( left ) or CD8 + ( middle ), from MRL/ lpr mice. Right , overlay histogram comparing the expression levels of P2X7R on B220 + DN T cells (–) and B220 – CD4 + T cells (–) from MRL/ lpr mice. The results shown are representative of at least four independent experiments.

    Techniques Used: Expressing, Flow Cytometry, Purification, Quantitative RT-PCR, Amplification, Western Blot, Affinity Purification, Staining, Generated

    rabbit anti p2x7r monoclonal igg  (Alomone Labs)


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    Structured Review

    Alomone Labs rabbit anti p2x7r monoclonal igg
    The sequences of oligonucleotide primers.
    Rabbit Anti P2x7r Monoclonal Igg, 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
    https://www.bioz.com/result/rabbit anti p2x7r monoclonal igg/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti p2x7r monoclonal igg - by Bioz Stars, 2023-03
    93/100 stars

    Images

    1) Product Images from "The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain"

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    Journal: BioMed Research International

    doi: 10.1155/2020/8143754

    The sequences of oligonucleotide primers.
    Figure Legend Snippet: The sequences of oligonucleotide primers.

    Techniques Used:

    The antibodies for immunoblotting.
    Figure Legend Snippet: The antibodies for immunoblotting.

    Techniques Used: Western Blot, Concentration Assay

    The antibodies for fluorescence labeling.
    Figure Legend Snippet: The antibodies for fluorescence labeling.

    Techniques Used: Fluorescence, Labeling, Concentration Assay

    Mechanical threshold and P2X7R and p-p38 expression. (a) Mechanical threshold after BTZ injection. (b, c) Western blot for P2X7R expression after BTZ treatment. (d) Immunofluorescence location of P2X7R in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. P2X7R is not expressed in NF-200-positive neurons. P2X7R is expressed in GFAP-labeled satellite glial cells (SGCs). (e) Immunofluorescence location of p-p38 in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. p-p38 is expressed in both MAP2-labeled neurons and GFAP-labeled SGCs. (f) Immunofluorescence location of P2X7R in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. P2X7R is expressed mainly in Iba-1-labeled microglial cells rather than in GFAP-labeled astrocytes and MAP2-labeled neurons. (g) Immunofluorescence location of p-p38 in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. p-p38 is expressed mainly in Iba-1-labeled microglial cells. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: Mechanical threshold and P2X7R and p-p38 expression. (a) Mechanical threshold after BTZ injection. (b, c) Western blot for P2X7R expression after BTZ treatment. (d) Immunofluorescence location of P2X7R in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. P2X7R is not expressed in NF-200-positive neurons. P2X7R is expressed in GFAP-labeled satellite glial cells (SGCs). (e) Immunofluorescence location of p-p38 in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. p-p38 is expressed in both MAP2-labeled neurons and GFAP-labeled SGCs. (f) Immunofluorescence location of P2X7R in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. P2X7R is expressed mainly in Iba-1-labeled microglial cells rather than in GFAP-labeled astrocytes and MAP2-labeled neurons. (g) Immunofluorescence location of p-p38 in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. p-p38 is expressed mainly in Iba-1-labeled microglial cells. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Injection, Western Blot, Immunofluorescence, Labeling

    p38 mRNA expression and p38 phosphorylation in DRG after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) p-p38 immunofluorescence labeling. The arrows show the typical p-p38 single-labeled DRG cells. (e) p-p38 fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗∗ P < 0.001.
    Figure Legend Snippet: p38 mRNA expression and p38 phosphorylation in DRG after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) p-p38 immunofluorescence labeling. The arrows show the typical p-p38 single-labeled DRG cells. (e) p-p38 fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Inhibition, Western Blot, Immunofluorescence, Labeling, Fluorescence

    p38 mRNA expression and p38 phosphorylation in SDH after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: p38 mRNA expression and p38 phosphorylation in SDH after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Inhibition, Western Blot, Fluorescence, Labeling

    IL-1 β , IL-6, and TNF- α mRNA expression in DRG and SDH after inhibition of P2X7R. (a) DRG IL-1 β mRNA. (b) DRG IL-6 mRNA. (c) DRG TNF- α mRNA. (d) SDH IL-1 β mRNA. (e) SDH IL-6 mRNA. (f) SDH TNF- α mRNA. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: IL-1 β , IL-6, and TNF- α mRNA expression in DRG and SDH after inhibition of P2X7R. (a) DRG IL-1 β mRNA. (b) DRG IL-6 mRNA. (c) DRG TNF- α mRNA. (d) SDH IL-1 β mRNA. (e) SDH IL-6 mRNA. (f) SDH TNF- α mRNA. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Inhibition

    P2X7R mRNA and protein expression in DRG after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and GFAP coexpression fluorescence labeling for SGCs. The arrows indicate the typical single-labeled and double-labeled DRG satellite cells. (e) P2X7R and GFAP coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: P2X7R mRNA and protein expression in DRG after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and GFAP coexpression fluorescence labeling for SGCs. The arrows indicate the typical single-labeled and double-labeled DRG satellite cells. (e) P2X7R and GFAP coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Inhibition, Western Blot, Fluorescence, Labeling

    P2X7R mRNA and protein expression in SDH after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 colocalization fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: P2X7R mRNA and protein expression in SDH after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 colocalization fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Inhibition, Western Blot, Fluorescence, Labeling

    Mechanical threshold alterations after inhibition of P2X7R or p38. (a) Mechanical threshold after inhibition of P2X7R. (b) Mechanical threshold after inhibition of p38. Mean ± SEM ( n = 5). ∗∗∗ P < 0.001 (vs. control); # P < 0.05; ## P < 0.01 (vs. BTZ group).
    Figure Legend Snippet: Mechanical threshold alterations after inhibition of P2X7R or p38. (a) Mechanical threshold after inhibition of P2X7R. (b) Mechanical threshold after inhibition of p38. Mean ± SEM ( n = 5). ∗∗∗ P < 0.001 (vs. control); # P < 0.05; ## P < 0.01 (vs. BTZ group).

    Techniques Used: Inhibition

    rabbit anti p2x7r monoclonal igg  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
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    Structured Review

    Alomone Labs rabbit anti p2x7r monoclonal igg
    The sequences of oligonucleotide primers.
    Rabbit Anti P2x7r Monoclonal Igg, 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
    https://www.bioz.com/result/rabbit anti p2x7r monoclonal igg/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti p2x7r monoclonal igg - by Bioz Stars, 2023-03
    93/100 stars

    Images

    1) Product Images from "The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain"

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    Journal: BioMed Research International

    doi: 10.1155/2020/8143754

    The sequences of oligonucleotide primers.
    Figure Legend Snippet: The sequences of oligonucleotide primers.

    Techniques Used:

    The antibodies for immunoblotting.
    Figure Legend Snippet: The antibodies for immunoblotting.

    Techniques Used: Western Blot, Concentration Assay

    The antibodies for fluorescence labeling.
    Figure Legend Snippet: The antibodies for fluorescence labeling.

    Techniques Used: Fluorescence, Labeling, Concentration Assay

    Mechanical threshold and P2X7R and p-p38 expression. (a) Mechanical threshold after BTZ injection. (b, c) Western blot for P2X7R expression after BTZ treatment. (d) Immunofluorescence location of P2X7R in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. P2X7R is not expressed in NF-200-positive neurons. P2X7R is expressed in GFAP-labeled satellite glial cells (SGCs). (e) Immunofluorescence location of p-p38 in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. p-p38 is expressed in both MAP2-labeled neurons and GFAP-labeled SGCs. (f) Immunofluorescence location of P2X7R in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. P2X7R is expressed mainly in Iba-1-labeled microglial cells rather than in GFAP-labeled astrocytes and MAP2-labeled neurons. (g) Immunofluorescence location of p-p38 in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. p-p38 is expressed mainly in Iba-1-labeled microglial cells. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: Mechanical threshold and P2X7R and p-p38 expression. (a) Mechanical threshold after BTZ injection. (b, c) Western blot for P2X7R expression after BTZ treatment. (d) Immunofluorescence location of P2X7R in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. P2X7R is not expressed in NF-200-positive neurons. P2X7R is expressed in GFAP-labeled satellite glial cells (SGCs). (e) Immunofluorescence location of p-p38 in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. p-p38 is expressed in both MAP2-labeled neurons and GFAP-labeled SGCs. (f) Immunofluorescence location of P2X7R in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. P2X7R is expressed mainly in Iba-1-labeled microglial cells rather than in GFAP-labeled astrocytes and MAP2-labeled neurons. (g) Immunofluorescence location of p-p38 in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. p-p38 is expressed mainly in Iba-1-labeled microglial cells. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Injection, Western Blot, Immunofluorescence, Labeling

    p38 mRNA expression and p38 phosphorylation in DRG after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) p-p38 immunofluorescence labeling. The arrows show the typical p-p38 single-labeled DRG cells. (e) p-p38 fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗∗ P < 0.001.
    Figure Legend Snippet: p38 mRNA expression and p38 phosphorylation in DRG after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) p-p38 immunofluorescence labeling. The arrows show the typical p-p38 single-labeled DRG cells. (e) p-p38 fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Inhibition, Western Blot, Immunofluorescence, Labeling, Fluorescence

    p38 mRNA expression and p38 phosphorylation in SDH after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: p38 mRNA expression and p38 phosphorylation in SDH after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Inhibition, Western Blot, Fluorescence, Labeling

    IL-1 β , IL-6, and TNF- α mRNA expression in DRG and SDH after inhibition of P2X7R. (a) DRG IL-1 β mRNA. (b) DRG IL-6 mRNA. (c) DRG TNF- α mRNA. (d) SDH IL-1 β mRNA. (e) SDH IL-6 mRNA. (f) SDH TNF- α mRNA. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: IL-1 β , IL-6, and TNF- α mRNA expression in DRG and SDH after inhibition of P2X7R. (a) DRG IL-1 β mRNA. (b) DRG IL-6 mRNA. (c) DRG TNF- α mRNA. (d) SDH IL-1 β mRNA. (e) SDH IL-6 mRNA. (f) SDH TNF- α mRNA. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Inhibition

    P2X7R mRNA and protein expression in DRG after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and GFAP coexpression fluorescence labeling for SGCs. The arrows indicate the typical single-labeled and double-labeled DRG satellite cells. (e) P2X7R and GFAP coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: P2X7R mRNA and protein expression in DRG after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and GFAP coexpression fluorescence labeling for SGCs. The arrows indicate the typical single-labeled and double-labeled DRG satellite cells. (e) P2X7R and GFAP coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Inhibition, Western Blot, Fluorescence, Labeling

    P2X7R mRNA and protein expression in SDH after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 colocalization fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: P2X7R mRNA and protein expression in SDH after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 colocalization fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Inhibition, Western Blot, Fluorescence, Labeling

    Mechanical threshold alterations after inhibition of P2X7R or p38. (a) Mechanical threshold after inhibition of P2X7R. (b) Mechanical threshold after inhibition of p38. Mean ± SEM ( n = 5). ∗∗∗ P < 0.001 (vs. control); # P < 0.05; ## P < 0.01 (vs. BTZ group).
    Figure Legend Snippet: Mechanical threshold alterations after inhibition of P2X7R or p38. (a) Mechanical threshold after inhibition of P2X7R. (b) Mechanical threshold after inhibition of p38. Mean ± SEM ( n = 5). ∗∗∗ P < 0.001 (vs. control); # P < 0.05; ## P < 0.01 (vs. BTZ group).

    Techniques Used: Inhibition

    rabbit p2x7r antibody  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
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    Structured Review

    Alomone Labs rabbit p2x7r antibody
    Immunofluorescence detected <t>P2X7R</t> of microglia in spinal cord. ( A ) P2X7R (P2X7R+, green) and IBA-1 (IBA-1+, red) co-expressed as P2X7R of microglia (P2X7R+/IBA-1+, yellow) that were overexpressed after spinal cord injury and BzATP intervention, and, compared to the morphology of microglia in the sham group, the morphology of activated microglia after injury changed to large soma and short irregular processes, revealing that many microglia in spinal cord became active after spinal cord injury. ( B ) The number of P2X7R of microglia (P2X7R+/IBA-1+) increased notably after spinal cord injury compared to the sham group (** P<0.01), and, compared to the model group, BzATP increased the number of P2X7R of microglia (* P<0.05) while A-438079 significantly decreased the number of P2X7R of microglia (** P<0.01). ( C ) The P2X7R-positive microglia percentage was clearly increased after spinal cord injury compared to the sham group (** P<0.01), and, compared to the model group, BzATP accounted for a significantly increased P2X7R-positive microglia proportion (** P<0.01) while A-438079 accounted for a significantly reduced P2X7R-positive microglia proportion (** P<0.01).
    Rabbit P2x7r Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit p2x7r antibody/product/Alomone Labs
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit p2x7r antibody - by Bioz Stars, 2023-03
    96/100 stars

    Images

    1) Product Images from "P2X7 Receptor (P2X7R) of Microglia Mediates Neuroinflammation by Regulating (NOD)-Like Receptor Protein 3 (NLRP3) Inflammasome-Dependent Inflammation After Spinal Cord Injury"

    Article Title: P2X7 Receptor (P2X7R) of Microglia Mediates Neuroinflammation by Regulating (NOD)-Like Receptor Protein 3 (NLRP3) Inflammasome-Dependent Inflammation After Spinal Cord Injury

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    doi: 10.12659/MSM.925491

    Immunofluorescence detected P2X7R of microglia in spinal cord. ( A ) P2X7R (P2X7R+, green) and IBA-1 (IBA-1+, red) co-expressed as P2X7R of microglia (P2X7R+/IBA-1+, yellow) that were overexpressed after spinal cord injury and BzATP intervention, and, compared to the morphology of microglia in the sham group, the morphology of activated microglia after injury changed to large soma and short irregular processes, revealing that many microglia in spinal cord became active after spinal cord injury. ( B ) The number of P2X7R of microglia (P2X7R+/IBA-1+) increased notably after spinal cord injury compared to the sham group (** P<0.01), and, compared to the model group, BzATP increased the number of P2X7R of microglia (* P<0.05) while A-438079 significantly decreased the number of P2X7R of microglia (** P<0.01). ( C ) The P2X7R-positive microglia percentage was clearly increased after spinal cord injury compared to the sham group (** P<0.01), and, compared to the model group, BzATP accounted for a significantly increased P2X7R-positive microglia proportion (** P<0.01) while A-438079 accounted for a significantly reduced P2X7R-positive microglia proportion (** P<0.01).
    Figure Legend Snippet: Immunofluorescence detected P2X7R of microglia in spinal cord. ( A ) P2X7R (P2X7R+, green) and IBA-1 (IBA-1+, red) co-expressed as P2X7R of microglia (P2X7R+/IBA-1+, yellow) that were overexpressed after spinal cord injury and BzATP intervention, and, compared to the morphology of microglia in the sham group, the morphology of activated microglia after injury changed to large soma and short irregular processes, revealing that many microglia in spinal cord became active after spinal cord injury. ( B ) The number of P2X7R of microglia (P2X7R+/IBA-1+) increased notably after spinal cord injury compared to the sham group (** P<0.01), and, compared to the model group, BzATP increased the number of P2X7R of microglia (* P<0.05) while A-438079 significantly decreased the number of P2X7R of microglia (** P<0.01). ( C ) The P2X7R-positive microglia percentage was clearly increased after spinal cord injury compared to the sham group (** P<0.01), and, compared to the model group, BzATP accounted for a significantly increased P2X7R-positive microglia proportion (** P<0.01) while A-438079 accounted for a significantly reduced P2X7R-positive microglia proportion (** P<0.01).

    Techniques Used: Immunofluorescence

    Western blot and qPCR assay were used to assess the expression of P2X7R protein and P2X7R mRNA in spinal cord. ( A ) Western blot assay showed that the expression of P2X7R protein in spinal cord was increased notably after spinal cord injury compared to the sham group (** P<0.01) and, compared to model group, BzATP increased the expression of P2X7R protein in spinal cord (* P<0.05) while A-438079 significantly decreased the expression of P2X7R protein in spinal cord (** P<0.01). ( B ) qPCR assay showed that the expression of P2X7R mRNA in spinal cord increased notably after spinal cord injury compared to the sham group (** P<0.01), and compared to the model group, BzATP increased the expression of P2X7R mRNA in spinal cord (* P<0.05) while A-438079 significantly decreased the expression of P2X7R mRNA in spinal cord (** P<0.01).
    Figure Legend Snippet: Western blot and qPCR assay were used to assess the expression of P2X7R protein and P2X7R mRNA in spinal cord. ( A ) Western blot assay showed that the expression of P2X7R protein in spinal cord was increased notably after spinal cord injury compared to the sham group (** P<0.01) and, compared to model group, BzATP increased the expression of P2X7R protein in spinal cord (* P<0.05) while A-438079 significantly decreased the expression of P2X7R protein in spinal cord (** P<0.01). ( B ) qPCR assay showed that the expression of P2X7R mRNA in spinal cord increased notably after spinal cord injury compared to the sham group (** P<0.01), and compared to the model group, BzATP increased the expression of P2X7R mRNA in spinal cord (* P<0.05) while A-438079 significantly decreased the expression of P2X7R mRNA in spinal cord (** P<0.01).

    Techniques Used: Western Blot, Expressing

    P2X7R of microglia in spinal cord-mediated neuroinflammation after spinal cord injury. ( A ) ELISA showed that, compared to the sham group, the expression of IL-1β and IL-18 in the spinal cord were significantly increased after spinal cord injury (** P<0.01) and, compared to the model group, A438079 significantly inhibited the expression of IL- 1β and IL-18 (** P<0.01) and BzATP significantly increased the expression of IL- 1β and IL-18 (** P<0.01), which was strongly inhibited by CY-09 (** P<0.01). ( B ) qPCR assay showed that, compared to the sham group, the expression of IL-1β mRNA and IL-18 mRNA in the spinal cord was significantly increased after spinal cord injury (** P<0.01) and, compared to the model group, A438079 significantly inhibited the expression of IL-1β mRNA and IL-18 mRNA (** P<0.01), and BzATP significantly increased the expression of IL-1β mRNA and IL-18 mRNA (** P<0.01), which was strongly inhibited by CY-09 (** P<0.01).
    Figure Legend Snippet: P2X7R of microglia in spinal cord-mediated neuroinflammation after spinal cord injury. ( A ) ELISA showed that, compared to the sham group, the expression of IL-1β and IL-18 in the spinal cord were significantly increased after spinal cord injury (** P<0.01) and, compared to the model group, A438079 significantly inhibited the expression of IL- 1β and IL-18 (** P<0.01) and BzATP significantly increased the expression of IL- 1β and IL-18 (** P<0.01), which was strongly inhibited by CY-09 (** P<0.01). ( B ) qPCR assay showed that, compared to the sham group, the expression of IL-1β mRNA and IL-18 mRNA in the spinal cord was significantly increased after spinal cord injury (** P<0.01) and, compared to the model group, A438079 significantly inhibited the expression of IL-1β mRNA and IL-18 mRNA (** P<0.01), and BzATP significantly increased the expression of IL-1β mRNA and IL-18 mRNA (** P<0.01), which was strongly inhibited by CY-09 (** P<0.01).

    Techniques Used: Enzyme-linked Immunosorbent Assay, Expressing

    P2X7R of microglia in spinal cord regulated the expression of NLRP3 and cleaved-Caspase-1 (P20). Compared to the sham group, the expression of NLRP3 and cleaved-Caspase-1 (P20) in the spinal cord significantly increased after spinal cord injury (** P<0.01) and, compared to the model group, BzATP promoted the expression of NLRP3 and cleaved-Caspase-1 (P20) (* P<0.05), which was inhibited by CY-09 (* P<0.05), while A-438079 notably reduced the expression of NLRP3 and cleaved-Caspase-1 (P20) (** P<0.01).
    Figure Legend Snippet: P2X7R of microglia in spinal cord regulated the expression of NLRP3 and cleaved-Caspase-1 (P20). Compared to the sham group, the expression of NLRP3 and cleaved-Caspase-1 (P20) in the spinal cord significantly increased after spinal cord injury (** P<0.01) and, compared to the model group, BzATP promoted the expression of NLRP3 and cleaved-Caspase-1 (P20) (* P<0.05), which was inhibited by CY-09 (* P<0.05), while A-438079 notably reduced the expression of NLRP3 and cleaved-Caspase-1 (P20) (** P<0.01).

    Techniques Used: Expressing

    rabbit p2x7r antibody  (Alomone Labs)


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    Alomone Labs rabbit p2x7r antibody
    Immunofluorescence detected <t>P2X7R</t> of microglia in spinal cord. ( A ) P2X7R (P2X7R+, green) and IBA-1 (IBA-1+, red) co-expressed as P2X7R of microglia (P2X7R+/IBA-1+, yellow) that were overexpressed after spinal cord injury and BzATP intervention, and, compared to the morphology of microglia in the sham group, the morphology of activated microglia after injury changed to large soma and short irregular processes, revealing that many microglia in spinal cord became active after spinal cord injury. ( B ) The number of P2X7R of microglia (P2X7R+/IBA-1+) increased notably after spinal cord injury compared to the sham group (** P<0.01), and, compared to the model group, BzATP increased the number of P2X7R of microglia (* P<0.05) while A-438079 significantly decreased the number of P2X7R of microglia (** P<0.01). ( C ) The P2X7R-positive microglia percentage was clearly increased after spinal cord injury compared to the sham group (** P<0.01), and, compared to the model group, BzATP accounted for a significantly increased P2X7R-positive microglia proportion (** P<0.01) while A-438079 accounted for a significantly reduced P2X7R-positive microglia proportion (** P<0.01).
    Rabbit P2x7r Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit p2x7r antibody/product/Alomone Labs
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit p2x7r antibody - by Bioz Stars, 2023-03
    96/100 stars

    Images

    1) Product Images from "P2X7 Receptor (P2X7R) of Microglia Mediates Neuroinflammation by Regulating (NOD)-Like Receptor Protein 3 (NLRP3) Inflammasome-Dependent Inflammation After Spinal Cord Injury"

    Article Title: P2X7 Receptor (P2X7R) of Microglia Mediates Neuroinflammation by Regulating (NOD)-Like Receptor Protein 3 (NLRP3) Inflammasome-Dependent Inflammation After Spinal Cord Injury

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    doi: 10.12659/MSM.925491

    Immunofluorescence detected P2X7R of microglia in spinal cord. ( A ) P2X7R (P2X7R+, green) and IBA-1 (IBA-1+, red) co-expressed as P2X7R of microglia (P2X7R+/IBA-1+, yellow) that were overexpressed after spinal cord injury and BzATP intervention, and, compared to the morphology of microglia in the sham group, the morphology of activated microglia after injury changed to large soma and short irregular processes, revealing that many microglia in spinal cord became active after spinal cord injury. ( B ) The number of P2X7R of microglia (P2X7R+/IBA-1+) increased notably after spinal cord injury compared to the sham group (** P<0.01), and, compared to the model group, BzATP increased the number of P2X7R of microglia (* P<0.05) while A-438079 significantly decreased the number of P2X7R of microglia (** P<0.01). ( C ) The P2X7R-positive microglia percentage was clearly increased after spinal cord injury compared to the sham group (** P<0.01), and, compared to the model group, BzATP accounted for a significantly increased P2X7R-positive microglia proportion (** P<0.01) while A-438079 accounted for a significantly reduced P2X7R-positive microglia proportion (** P<0.01).
    Figure Legend Snippet: Immunofluorescence detected P2X7R of microglia in spinal cord. ( A ) P2X7R (P2X7R+, green) and IBA-1 (IBA-1+, red) co-expressed as P2X7R of microglia (P2X7R+/IBA-1+, yellow) that were overexpressed after spinal cord injury and BzATP intervention, and, compared to the morphology of microglia in the sham group, the morphology of activated microglia after injury changed to large soma and short irregular processes, revealing that many microglia in spinal cord became active after spinal cord injury. ( B ) The number of P2X7R of microglia (P2X7R+/IBA-1+) increased notably after spinal cord injury compared to the sham group (** P<0.01), and, compared to the model group, BzATP increased the number of P2X7R of microglia (* P<0.05) while A-438079 significantly decreased the number of P2X7R of microglia (** P<0.01). ( C ) The P2X7R-positive microglia percentage was clearly increased after spinal cord injury compared to the sham group (** P<0.01), and, compared to the model group, BzATP accounted for a significantly increased P2X7R-positive microglia proportion (** P<0.01) while A-438079 accounted for a significantly reduced P2X7R-positive microglia proportion (** P<0.01).

    Techniques Used: Immunofluorescence

    Western blot and qPCR assay were used to assess the expression of P2X7R protein and P2X7R mRNA in spinal cord. ( A ) Western blot assay showed that the expression of P2X7R protein in spinal cord was increased notably after spinal cord injury compared to the sham group (** P<0.01) and, compared to model group, BzATP increased the expression of P2X7R protein in spinal cord (* P<0.05) while A-438079 significantly decreased the expression of P2X7R protein in spinal cord (** P<0.01). ( B ) qPCR assay showed that the expression of P2X7R mRNA in spinal cord increased notably after spinal cord injury compared to the sham group (** P<0.01), and compared to the model group, BzATP increased the expression of P2X7R mRNA in spinal cord (* P<0.05) while A-438079 significantly decreased the expression of P2X7R mRNA in spinal cord (** P<0.01).
    Figure Legend Snippet: Western blot and qPCR assay were used to assess the expression of P2X7R protein and P2X7R mRNA in spinal cord. ( A ) Western blot assay showed that the expression of P2X7R protein in spinal cord was increased notably after spinal cord injury compared to the sham group (** P<0.01) and, compared to model group, BzATP increased the expression of P2X7R protein in spinal cord (* P<0.05) while A-438079 significantly decreased the expression of P2X7R protein in spinal cord (** P<0.01). ( B ) qPCR assay showed that the expression of P2X7R mRNA in spinal cord increased notably after spinal cord injury compared to the sham group (** P<0.01), and compared to the model group, BzATP increased the expression of P2X7R mRNA in spinal cord (* P<0.05) while A-438079 significantly decreased the expression of P2X7R mRNA in spinal cord (** P<0.01).

    Techniques Used: Western Blot, Expressing

    P2X7R of microglia in spinal cord-mediated neuroinflammation after spinal cord injury. ( A ) ELISA showed that, compared to the sham group, the expression of IL-1β and IL-18 in the spinal cord were significantly increased after spinal cord injury (** P<0.01) and, compared to the model group, A438079 significantly inhibited the expression of IL- 1β and IL-18 (** P<0.01) and BzATP significantly increased the expression of IL- 1β and IL-18 (** P<0.01), which was strongly inhibited by CY-09 (** P<0.01). ( B ) qPCR assay showed that, compared to the sham group, the expression of IL-1β mRNA and IL-18 mRNA in the spinal cord was significantly increased after spinal cord injury (** P<0.01) and, compared to the model group, A438079 significantly inhibited the expression of IL-1β mRNA and IL-18 mRNA (** P<0.01), and BzATP significantly increased the expression of IL-1β mRNA and IL-18 mRNA (** P<0.01), which was strongly inhibited by CY-09 (** P<0.01).
    Figure Legend Snippet: P2X7R of microglia in spinal cord-mediated neuroinflammation after spinal cord injury. ( A ) ELISA showed that, compared to the sham group, the expression of IL-1β and IL-18 in the spinal cord were significantly increased after spinal cord injury (** P<0.01) and, compared to the model group, A438079 significantly inhibited the expression of IL- 1β and IL-18 (** P<0.01) and BzATP significantly increased the expression of IL- 1β and IL-18 (** P<0.01), which was strongly inhibited by CY-09 (** P<0.01). ( B ) qPCR assay showed that, compared to the sham group, the expression of IL-1β mRNA and IL-18 mRNA in the spinal cord was significantly increased after spinal cord injury (** P<0.01) and, compared to the model group, A438079 significantly inhibited the expression of IL-1β mRNA and IL-18 mRNA (** P<0.01), and BzATP significantly increased the expression of IL-1β mRNA and IL-18 mRNA (** P<0.01), which was strongly inhibited by CY-09 (** P<0.01).

    Techniques Used: Enzyme-linked Immunosorbent Assay, Expressing

    P2X7R of microglia in spinal cord regulated the expression of NLRP3 and cleaved-Caspase-1 (P20). Compared to the sham group, the expression of NLRP3 and cleaved-Caspase-1 (P20) in the spinal cord significantly increased after spinal cord injury (** P<0.01) and, compared to the model group, BzATP promoted the expression of NLRP3 and cleaved-Caspase-1 (P20) (* P<0.05), which was inhibited by CY-09 (* P<0.05), while A-438079 notably reduced the expression of NLRP3 and cleaved-Caspase-1 (P20) (** P<0.01).
    Figure Legend Snippet: P2X7R of microglia in spinal cord regulated the expression of NLRP3 and cleaved-Caspase-1 (P20). Compared to the sham group, the expression of NLRP3 and cleaved-Caspase-1 (P20) in the spinal cord significantly increased after spinal cord injury (** P<0.01) and, compared to the model group, BzATP promoted the expression of NLRP3 and cleaved-Caspase-1 (P20) (* P<0.05), which was inhibited by CY-09 (* P<0.05), while A-438079 notably reduced the expression of NLRP3 and cleaved-Caspase-1 (P20) (** P<0.01).

    Techniques Used: Expressing

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    • anti p2x7r  (Alomone Labs)
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    Alomone Labs anti p2x7r
    Primary antibodies used for Western Blot (WB) and immunohistochemistry (IHC).
    Anti P2x7r, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 96 stars, based on 1 article reviews
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    p2x7r staining  (Alomone Labs)
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    Alomone Labs p2x7r staining
    <t>P2X7R</t> is present in neurons cultivated in vitro during development. A , Scheme of the experiment. B , Scheme of the regions amplified by qPCR. C , Analysis of the expression of extracellular and ( D ) intracellular sequences of P2X7R by real-time qPCR (DIV1) on each DIV. Relative expression is presented as the fold change normalized to GAPDH expression. E , WT cells were cultured for the indicated period of time, and P2X7R protein expression was determined by immunoblotting. Graphs represent densitometric evaluation of the target protein band normalized to the total amount of loaded protein ( n = 4). F , To validate the specificity of the P2X7R antibody, WT and P2X7R-KO cells were cultured for the indicated period of time, and P2X7R protein expression was determined by immunoblotting. Graphs represent the densitometric evaluation of the target protein band normalized to total amount of loaded protein ( n = 4). G , Areas under the curve for WT and KO primary pyramidal cells at DIV4 after acute BzATP (1 m m ) application. H , Areas under the curve for WT and KO primary pyramidal cells at DIV4 after acute calcium ionophore (1 m m ) application. I , Immunostaining of a neuronal marker (MAP2) and P2X7R in KO and WT primary neurons at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 100 µm. J , Immunostaining for a transfection marker (eGFP) and P2X7R at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 200 µm. * p < 0.05. ** p < 0.005. **** p < 0.0001. ns, non-significant. Data presented as Mean±SEM.
    P2x7r Staining, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    p2x7r  (Alomone Labs)
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    Alomone Labs p2x7r
    Sequences of siRNA to <t> P2X7R. </t>
    P2x7r, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    affinity purified rabbit anti p2x7r antibodies  (Alomone Labs)
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    Alomone Labs affinity purified rabbit anti p2x7r antibodies
    Spleen cells from 3 to 4-mo-old MRL +/+ ( solid line ), MRL/ lpr ( dashed line ) and P2X7-deficient B6 <t>P2X7R−/−</t> ( dotted line ) mice (n = 3 mice/group) were treated for 45 min at 37°C with doses of ATP ranging from 100 to 5000 µM. Spleen cells were then triple-stained with anti-CD90, anti-CD19 and anti-CD62L mAb to assess by flow cytometry: (A) the percentage of CD62L-expressing CD19 – CD90 + T cells and (B) MFI of CD62L on CD19 – CD90 + T cells. Results are expressed as the mean percentage of initial expression ± SE.
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    rabbit anti p2x7r monoclonal igg  (Alomone Labs)
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    Alomone Labs rabbit anti p2x7r monoclonal igg
    The sequences of oligonucleotide primers.
    Rabbit Anti P2x7r Monoclonal Igg, 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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    rabbit p2x7r antibody  (Alomone Labs)
    96
    Alomone Labs rabbit p2x7r antibody
    Immunofluorescence detected <t>P2X7R</t> of microglia in spinal cord. ( A ) P2X7R (P2X7R+, green) and IBA-1 (IBA-1+, red) co-expressed as P2X7R of microglia (P2X7R+/IBA-1+, yellow) that were overexpressed after spinal cord injury and BzATP intervention, and, compared to the morphology of microglia in the sham group, the morphology of activated microglia after injury changed to large soma and short irregular processes, revealing that many microglia in spinal cord became active after spinal cord injury. ( B ) The number of P2X7R of microglia (P2X7R+/IBA-1+) increased notably after spinal cord injury compared to the sham group (** P<0.01), and, compared to the model group, BzATP increased the number of P2X7R of microglia (* P<0.05) while A-438079 significantly decreased the number of P2X7R of microglia (** P<0.01). ( C ) The P2X7R-positive microglia percentage was clearly increased after spinal cord injury compared to the sham group (** P<0.01), and, compared to the model group, BzATP accounted for a significantly increased P2X7R-positive microglia proportion (** P<0.01) while A-438079 accounted for a significantly reduced P2X7R-positive microglia proportion (** P<0.01).
    Rabbit P2x7r Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit p2x7r antibody/product/Alomone Labs
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    rabbit p2x7r antibody - by Bioz Stars, 2023-03
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    Image Search Results


    Primary antibodies used for Western Blot (WB) and immunohistochemistry (IHC).

    Journal: PLoS ONE

    Article Title: A Co-Culture System with an Organotypic Lung Slice and an Immortal Alveolar Macrophage Cell Line to Quantify Silica-Induced Inflammation

    doi: 10.1371/journal.pone.0117056

    Figure Lengend Snippet: Primary antibodies used for Western Blot (WB) and immunohistochemistry (IHC).

    Article Snippet: anti-P2X7R , 1:500 , - , Rabbit , Polyclonal , Alomone Labs (Jerusalem, Israel).

    Techniques: Western Blot, Immunohistochemistry

    P2X7R is present in neurons cultivated in vitro during development. A , Scheme of the experiment. B , Scheme of the regions amplified by qPCR. C , Analysis of the expression of extracellular and ( D ) intracellular sequences of P2X7R by real-time qPCR (DIV1) on each DIV. Relative expression is presented as the fold change normalized to GAPDH expression. E , WT cells were cultured for the indicated period of time, and P2X7R protein expression was determined by immunoblotting. Graphs represent densitometric evaluation of the target protein band normalized to the total amount of loaded protein ( n = 4). F , To validate the specificity of the P2X7R antibody, WT and P2X7R-KO cells were cultured for the indicated period of time, and P2X7R protein expression was determined by immunoblotting. Graphs represent the densitometric evaluation of the target protein band normalized to total amount of loaded protein ( n = 4). G , Areas under the curve for WT and KO primary pyramidal cells at DIV4 after acute BzATP (1 m m ) application. H , Areas under the curve for WT and KO primary pyramidal cells at DIV4 after acute calcium ionophore (1 m m ) application. I , Immunostaining of a neuronal marker (MAP2) and P2X7R in KO and WT primary neurons at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 100 µm. J , Immunostaining for a transfection marker (eGFP) and P2X7R at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 200 µm. * p < 0.05. ** p < 0.005. **** p < 0.0001. ns, non-significant. Data presented as Mean±SEM.

    Journal: The Journal of Neuroscience

    Article Title: Dual Role of the P2X7 Receptor in Dendritic Outgrowth during Physiological and Pathological Brain Development

    doi: 10.1523/JNEUROSCI.0805-22.2022

    Figure Lengend Snippet: P2X7R is present in neurons cultivated in vitro during development. A , Scheme of the experiment. B , Scheme of the regions amplified by qPCR. C , Analysis of the expression of extracellular and ( D ) intracellular sequences of P2X7R by real-time qPCR (DIV1) on each DIV. Relative expression is presented as the fold change normalized to GAPDH expression. E , WT cells were cultured for the indicated period of time, and P2X7R protein expression was determined by immunoblotting. Graphs represent densitometric evaluation of the target protein band normalized to the total amount of loaded protein ( n = 4). F , To validate the specificity of the P2X7R antibody, WT and P2X7R-KO cells were cultured for the indicated period of time, and P2X7R protein expression was determined by immunoblotting. Graphs represent the densitometric evaluation of the target protein band normalized to total amount of loaded protein ( n = 4). G , Areas under the curve for WT and KO primary pyramidal cells at DIV4 after acute BzATP (1 m m ) application. H , Areas under the curve for WT and KO primary pyramidal cells at DIV4 after acute calcium ionophore (1 m m ) application. I , Immunostaining of a neuronal marker (MAP2) and P2X7R in KO and WT primary neurons at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 100 µm. J , Immunostaining for a transfection marker (eGFP) and P2X7R at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 200 µm. * p < 0.05. ** p < 0.005. **** p < 0.0001. ns, non-significant. Data presented as Mean±SEM.

    Article Snippet: For P2X7R staining, cells were incubated with a rabbit α-P2X7 antibody (Alomone Labs, RRID: AB_2040068 ) diluted 1:100 for two nights and then with AlexaFluor-549-conjugated anti-rabbit (Invitrogen, RRID: AB_2762824 ) and AlexaFluor-488-conjugated anti-rabbit (Invitrogen, RRID: AB_2633280 ) secondary antibodies diluted 1:500.

    Techniques: In Vitro, Amplification, Expressing, Cell Culture, Western Blot, Immunostaining, Marker, Transfection

    Neurons from P2rx7 −/− mice present less dendritic outgrowth. A , Scheme of the in vitro experiment. B , Immunostaining of a neuronal marker (MAP2) and an astroglial marker (GFAP) and of a transfection marker (eGFP) and a neuronal marker (MAP2) as the control for the transfection in a representative control WT primary hippocampal neuron at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 200 µm. C , Representative hippocampal neurons from WT and P2X7R KO mice transfected with and stained for eGFP. D , Sholl analysis showing the number of intersections versus distance from the soma (radius, μm). E , Using HPLC, nucleotide concentrations in the supernatants of WT and KO primary hippocampal neurons were measured. Quantification of the ( F ) cell body area, ( G ) number of primary dendrites, ( H ) number of endings, and ( I ) mean dendritic length. ** p < 0.005. **** p < 0.0001. ns, non-significant. Data presented as Mean±SEM.

    Journal: The Journal of Neuroscience

    Article Title: Dual Role of the P2X7 Receptor in Dendritic Outgrowth during Physiological and Pathological Brain Development

    doi: 10.1523/JNEUROSCI.0805-22.2022

    Figure Lengend Snippet: Neurons from P2rx7 −/− mice present less dendritic outgrowth. A , Scheme of the in vitro experiment. B , Immunostaining of a neuronal marker (MAP2) and an astroglial marker (GFAP) and of a transfection marker (eGFP) and a neuronal marker (MAP2) as the control for the transfection in a representative control WT primary hippocampal neuron at DIV10. Nuclei were counterstained with Hoechst. Scale bar, 200 µm. C , Representative hippocampal neurons from WT and P2X7R KO mice transfected with and stained for eGFP. D , Sholl analysis showing the number of intersections versus distance from the soma (radius, μm). E , Using HPLC, nucleotide concentrations in the supernatants of WT and KO primary hippocampal neurons were measured. Quantification of the ( F ) cell body area, ( G ) number of primary dendrites, ( H ) number of endings, and ( I ) mean dendritic length. ** p < 0.005. **** p < 0.0001. ns, non-significant. Data presented as Mean±SEM.

    Article Snippet: For P2X7R staining, cells were incubated with a rabbit α-P2X7 antibody (Alomone Labs, RRID: AB_2040068 ) diluted 1:100 for two nights and then with AlexaFluor-549-conjugated anti-rabbit (Invitrogen, RRID: AB_2762824 ) and AlexaFluor-488-conjugated anti-rabbit (Invitrogen, RRID: AB_2633280 ) secondary antibodies diluted 1:500.

    Techniques: In Vitro, Immunostaining, Marker, Transfection, Staining

    Fine-tuning effect of P2X7R on dendritic outgrowth. Antagonists disrupt normal dendritic outgrowth in vitro . A , Scheme of the in vitro experiment. B , Sholl analysis showing the number of intersections versus distance from the soma. Quantification of the ( C ) cell body area, ( D ) number of primary dendrites, ( E ) number of endings, and ( F ) mean dendritic length. Neurons acutely treated with ARL 67156 (100 U/ml) present elongation of dendritic endings. G , Scheme of the in vitro experiment. H , Sholl analysis showing the number of intersections versus distance from the soma. I , Using HPLC, nucleotide concentrations in the supernatants of WT and ARL 67156-treated WT primary hippocampal neurons were measured. Quantification of the ( J ) cell body area, ( K ) number of primary dendrites, ( L ) number of endings, and ( M ) mean total dendritic length. * p < 0.05. ** p < 0.005. *** p < 0.0005. **** p < 0.0001. ns, non-significant. Data presented as Mean±SEM.

    Journal: The Journal of Neuroscience

    Article Title: Dual Role of the P2X7 Receptor in Dendritic Outgrowth during Physiological and Pathological Brain Development

    doi: 10.1523/JNEUROSCI.0805-22.2022

    Figure Lengend Snippet: Fine-tuning effect of P2X7R on dendritic outgrowth. Antagonists disrupt normal dendritic outgrowth in vitro . A , Scheme of the in vitro experiment. B , Sholl analysis showing the number of intersections versus distance from the soma. Quantification of the ( C ) cell body area, ( D ) number of primary dendrites, ( E ) number of endings, and ( F ) mean dendritic length. Neurons acutely treated with ARL 67156 (100 U/ml) present elongation of dendritic endings. G , Scheme of the in vitro experiment. H , Sholl analysis showing the number of intersections versus distance from the soma. I , Using HPLC, nucleotide concentrations in the supernatants of WT and ARL 67156-treated WT primary hippocampal neurons were measured. Quantification of the ( J ) cell body area, ( K ) number of primary dendrites, ( L ) number of endings, and ( M ) mean total dendritic length. * p < 0.05. ** p < 0.005. *** p < 0.0005. **** p < 0.0001. ns, non-significant. Data presented as Mean±SEM.

    Article Snippet: For P2X7R staining, cells were incubated with a rabbit α-P2X7 antibody (Alomone Labs, RRID: AB_2040068 ) diluted 1:100 for two nights and then with AlexaFluor-549-conjugated anti-rabbit (Invitrogen, RRID: AB_2762824 ) and AlexaFluor-488-conjugated anti-rabbit (Invitrogen, RRID: AB_2633280 ) secondary antibodies diluted 1:500.

    Techniques: In Vitro

    Pyramidal neurons from P2rx7 −/− mice present less dendritic outgrowth in the CA1 region of the hippocampus. A , Representative pyramidal hippocampal neurons in slices of the hippocampal CA1 region from P20-29 WT and P2X7R KO mice. B , Sholl analysis showing the number of intersections versus distance from the soma for WT and KO pyramidal neurons. Quantification of the ( C ) cell body area, ( D ) number of primary dendrites, ( E ) number of endings, and ( F ) mean dendritic length. G , Representative pyramidal hippocampal neurons in slices of the hippocampal CA3 region from P20-29 WT and KO mice. H , Sholl analysis showing the number of intersections versus distance from the soma for WT and KO pyramidal neurons. Data are mean ± SEM and analyzed using two-way ANOVA followed by Bonferroni's multiple comparison test. Quantification of the ( I ) cell body area, ( J ) number of primary dendrites, ( K ) number of endings, and ( L ) mean dendritic length. Morphologic deficits correlated with observed deficits in cognitive performance in younger animals. Different aspects of episodic memory, such as ( M ) OL and ( N ) TOR, were analyzed. Deficits in cognitive performance were observed according to both ( O ) the OL index (%) and ( P ) the TOR index (%). Q , Scheme of the experiment. NOR was analyzed ( R ) 2 min after the exploration phase and ( S ) after a longer delay of 24 h. ** p < 0.005. **** p < 0.0001. * p < 0.05. ns, non-significant. Data presented as Mean±SEM.

    Journal: The Journal of Neuroscience

    Article Title: Dual Role of the P2X7 Receptor in Dendritic Outgrowth during Physiological and Pathological Brain Development

    doi: 10.1523/JNEUROSCI.0805-22.2022

    Figure Lengend Snippet: Pyramidal neurons from P2rx7 −/− mice present less dendritic outgrowth in the CA1 region of the hippocampus. A , Representative pyramidal hippocampal neurons in slices of the hippocampal CA1 region from P20-29 WT and P2X7R KO mice. B , Sholl analysis showing the number of intersections versus distance from the soma for WT and KO pyramidal neurons. Quantification of the ( C ) cell body area, ( D ) number of primary dendrites, ( E ) number of endings, and ( F ) mean dendritic length. G , Representative pyramidal hippocampal neurons in slices of the hippocampal CA3 region from P20-29 WT and KO mice. H , Sholl analysis showing the number of intersections versus distance from the soma for WT and KO pyramidal neurons. Data are mean ± SEM and analyzed using two-way ANOVA followed by Bonferroni's multiple comparison test. Quantification of the ( I ) cell body area, ( J ) number of primary dendrites, ( K ) number of endings, and ( L ) mean dendritic length. Morphologic deficits correlated with observed deficits in cognitive performance in younger animals. Different aspects of episodic memory, such as ( M ) OL and ( N ) TOR, were analyzed. Deficits in cognitive performance were observed according to both ( O ) the OL index (%) and ( P ) the TOR index (%). Q , Scheme of the experiment. NOR was analyzed ( R ) 2 min after the exploration phase and ( S ) after a longer delay of 24 h. ** p < 0.005. **** p < 0.0001. * p < 0.05. ns, non-significant. Data presented as Mean±SEM.

    Article Snippet: For P2X7R staining, cells were incubated with a rabbit α-P2X7 antibody (Alomone Labs, RRID: AB_2040068 ) diluted 1:100 for two nights and then with AlexaFluor-549-conjugated anti-rabbit (Invitrogen, RRID: AB_2762824 ) and AlexaFluor-488-conjugated anti-rabbit (Invitrogen, RRID: AB_2633280 ) secondary antibodies diluted 1:500.

    Techniques:

    P2X7R-deficient mice present weaker synaptic strength. A , Representative traces from the WT and KO groups in the presence of gabazine. B , AMPA/NMDA ratio. C , AMPA-mediated current amplitude. D , NMDA-mediated current amplitude. E , AMPA-mediated current rise time. F , AMPA-mediated current decay time. G , NMDA-mediated current rise time. H , NMDA-mediated current decay time. I , Number of spines on WT and KO neurons. J , Representative segments of WT and KO neurons used to count spines. Scale bar, 50 µm. ** p < 0.005. *** p < 0.0005. ns, non-significant. Data presented as Mean±SEM.

    Journal: The Journal of Neuroscience

    Article Title: Dual Role of the P2X7 Receptor in Dendritic Outgrowth during Physiological and Pathological Brain Development

    doi: 10.1523/JNEUROSCI.0805-22.2022

    Figure Lengend Snippet: P2X7R-deficient mice present weaker synaptic strength. A , Representative traces from the WT and KO groups in the presence of gabazine. B , AMPA/NMDA ratio. C , AMPA-mediated current amplitude. D , NMDA-mediated current amplitude. E , AMPA-mediated current rise time. F , AMPA-mediated current decay time. G , NMDA-mediated current rise time. H , NMDA-mediated current decay time. I , Number of spines on WT and KO neurons. J , Representative segments of WT and KO neurons used to count spines. Scale bar, 50 µm. ** p < 0.005. *** p < 0.0005. ns, non-significant. Data presented as Mean±SEM.

    Article Snippet: For P2X7R staining, cells were incubated with a rabbit α-P2X7 antibody (Alomone Labs, RRID: AB_2040068 ) diluted 1:100 for two nights and then with AlexaFluor-549-conjugated anti-rabbit (Invitrogen, RRID: AB_2762824 ) and AlexaFluor-488-conjugated anti-rabbit (Invitrogen, RRID: AB_2633280 ) secondary antibodies diluted 1:500.

    Techniques:

    P2X7R plays a role in a neurodevelopmental model of schizophrenia. A , B , Sholl analysis showing the number of intersections versus distance from the soma for WT neurons, WT neurons treated with different doses of saline or PIC on E12.5, and KO neurons. Quantification of the ( C ) cell body area, ( D ) number of primary dendrites, ( E ) number of endings, and ( F ) mean dendritic length for WT neurons after different treatments. Quantification of the ( G ) cell body area, ( H ) number of primary dendrites, ( I ) number of endings, and ( J ) mean dendritic length for KO neurons after different treatments. K , Using HPLC, nucleotide concentrations in the supernatants of WT and PIC-treated WT primary hippocampal neurons were measured. L , Using HPLC, nucleotide concentrations in the supernatants of KO- and PIC-treated KO primary hippocampal neurons were measured. *** p < 0.0005. **** p < 0.0001. Data presented as Mean±SEM.

    Journal: The Journal of Neuroscience

    Article Title: Dual Role of the P2X7 Receptor in Dendritic Outgrowth during Physiological and Pathological Brain Development

    doi: 10.1523/JNEUROSCI.0805-22.2022

    Figure Lengend Snippet: P2X7R plays a role in a neurodevelopmental model of schizophrenia. A , B , Sholl analysis showing the number of intersections versus distance from the soma for WT neurons, WT neurons treated with different doses of saline or PIC on E12.5, and KO neurons. Quantification of the ( C ) cell body area, ( D ) number of primary dendrites, ( E ) number of endings, and ( F ) mean dendritic length for WT neurons after different treatments. Quantification of the ( G ) cell body area, ( H ) number of primary dendrites, ( I ) number of endings, and ( J ) mean dendritic length for KO neurons after different treatments. K , Using HPLC, nucleotide concentrations in the supernatants of WT and PIC-treated WT primary hippocampal neurons were measured. L , Using HPLC, nucleotide concentrations in the supernatants of KO- and PIC-treated KO primary hippocampal neurons were measured. *** p < 0.0005. **** p < 0.0001. Data presented as Mean±SEM.

    Article Snippet: For P2X7R staining, cells were incubated with a rabbit α-P2X7 antibody (Alomone Labs, RRID: AB_2040068 ) diluted 1:100 for two nights and then with AlexaFluor-549-conjugated anti-rabbit (Invitrogen, RRID: AB_2762824 ) and AlexaFluor-488-conjugated anti-rabbit (Invitrogen, RRID: AB_2633280 ) secondary antibodies diluted 1:500.

    Techniques:

    P2X7R drives PIC-induced schizophrenic-like behavior in mice. A , Overview of the experimental protocol. B , Maternal PIC induced elevation in the level of IL-1β in fetal brain samples of WT mice, while this effect was not observed in samples from KO. Values measured are expressed in picograms per mg of total protein. C , Total distance moved (cm) and ( D ) velocity (cm/s) in the open field were not different between WT animals and PIC-treated WT animals. Quantification of ( E ) the spontaneous alternation percentage and ( F ) NOR percentage in the T-maze test showing that MIA-exposed WT animals exhibited cognitive deficits. G , The amounts of time spent exploring the different cages (seconds) in the social preference test were compared between animals of different genotypes and animal subjected to different treatments. H , The startle response was not different between WT and PIC-treated WT animals. I , PPI was disrupted in PIC-treated WT animals. * p < 0.05. ** p < 0.005. *** p < 0.0005. Data presented as Mean±SEM.

    Journal: The Journal of Neuroscience

    Article Title: Dual Role of the P2X7 Receptor in Dendritic Outgrowth during Physiological and Pathological Brain Development

    doi: 10.1523/JNEUROSCI.0805-22.2022

    Figure Lengend Snippet: P2X7R drives PIC-induced schizophrenic-like behavior in mice. A , Overview of the experimental protocol. B , Maternal PIC induced elevation in the level of IL-1β in fetal brain samples of WT mice, while this effect was not observed in samples from KO. Values measured are expressed in picograms per mg of total protein. C , Total distance moved (cm) and ( D ) velocity (cm/s) in the open field were not different between WT animals and PIC-treated WT animals. Quantification of ( E ) the spontaneous alternation percentage and ( F ) NOR percentage in the T-maze test showing that MIA-exposed WT animals exhibited cognitive deficits. G , The amounts of time spent exploring the different cages (seconds) in the social preference test were compared between animals of different genotypes and animal subjected to different treatments. H , The startle response was not different between WT and PIC-treated WT animals. I , PPI was disrupted in PIC-treated WT animals. * p < 0.05. ** p < 0.005. *** p < 0.0005. Data presented as Mean±SEM.

    Article Snippet: For P2X7R staining, cells were incubated with a rabbit α-P2X7 antibody (Alomone Labs, RRID: AB_2040068 ) diluted 1:100 for two nights and then with AlexaFluor-549-conjugated anti-rabbit (Invitrogen, RRID: AB_2762824 ) and AlexaFluor-488-conjugated anti-rabbit (Invitrogen, RRID: AB_2633280 ) secondary antibodies diluted 1:500.

    Techniques:

    Statistical analysis for molecular and morphologic analysis

    Journal: The Journal of Neuroscience

    Article Title: Dual Role of the P2X7 Receptor in Dendritic Outgrowth during Physiological and Pathological Brain Development

    doi: 10.1523/JNEUROSCI.0805-22.2022

    Figure Lengend Snippet: Statistical analysis for molecular and morphologic analysis

    Article Snippet: For P2X7R staining, cells were incubated with a rabbit α-P2X7 antibody (Alomone Labs, RRID: AB_2040068 ) diluted 1:100 for two nights and then with AlexaFluor-549-conjugated anti-rabbit (Invitrogen, RRID: AB_2762824 ) and AlexaFluor-488-conjugated anti-rabbit (Invitrogen, RRID: AB_2633280 ) secondary antibodies diluted 1:500.

    Techniques: Western Blot, Imaging, Enzyme-linked Immunosorbent Assay

    Sequences of siRNA to P2X7R.

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Inhibition of P2X7 Purinergic Receptor Ameliorates Cardiac Fibrosis by Suppressing NLRP3/IL-1 β Pathway

    doi: 10.1155/2020/7956274

    Figure Lengend Snippet: Sequences of siRNA to P2X7R.

    Article Snippet: The primary antibody for incubation was anti- α -SMA (Abcam, 1 : 200) or P2X7R (Alomone Labs, 1 : 200).

    Techniques:

    Rat RT-qPCR primer sequences.

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Inhibition of P2X7 Purinergic Receptor Ameliorates Cardiac Fibrosis by Suppressing NLRP3/IL-1 β Pathway

    doi: 10.1155/2020/7956274

    Figure Lengend Snippet: Rat RT-qPCR primer sequences.

    Article Snippet: The primary antibody for incubation was anti- α -SMA (Abcam, 1 : 200) or P2X7R (Alomone Labs, 1 : 200).

    Techniques: Sequencing

    Mice RT-qPCR primer sequences.

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Inhibition of P2X7 Purinergic Receptor Ameliorates Cardiac Fibrosis by Suppressing NLRP3/IL-1 β Pathway

    doi: 10.1155/2020/7956274

    Figure Lengend Snippet: Mice RT-qPCR primer sequences.

    Article Snippet: The primary antibody for incubation was anti- α -SMA (Abcam, 1 : 200) or P2X7R (Alomone Labs, 1 : 200).

    Techniques: Sequencing

    P2X7R was upregulated in the mice heart after TAC surgery. (a) Cardiac images and HW/BW ( n = 8). (b) HE-, Sirius Red-, and Masson's trichrome-stained sections of the hearts following four weeks of TAC treatment ( n = 6). (c) Representative images of an echocardiographic assessment of mice after TAC (4 weeks). Cardiac function indicators measured by echocardiography (LVEF (%), FS (%), LVPWd, IVSd, and LVIDd) in TAC- and sham-operated mice. (d) Expression of CTGF, periostin, and α -SMA mRNAs and proteins in heart cells ( n = 6). (e) Gene expression of P2X receptors in TAC- and sham-operated mice ( n = 6) at 4 weeks and gene expression of P2X7R in heart cells at 3 days, 1 week, 2 weeks, and 4 weeks after TAC- or sham-operated treatment ( n = 6). (f) Expression of P2X7R protein in mouse heart cells ( n = 6). Results are presented as means ± standard error of the mean. ∗ indicates P < 0.05, ∗∗ indicates P < 0.01, and ∗∗∗ indicates P < 0.001 versus sham.

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Inhibition of P2X7 Purinergic Receptor Ameliorates Cardiac Fibrosis by Suppressing NLRP3/IL-1 β Pathway

    doi: 10.1155/2020/7956274

    Figure Lengend Snippet: P2X7R was upregulated in the mice heart after TAC surgery. (a) Cardiac images and HW/BW ( n = 8). (b) HE-, Sirius Red-, and Masson's trichrome-stained sections of the hearts following four weeks of TAC treatment ( n = 6). (c) Representative images of an echocardiographic assessment of mice after TAC (4 weeks). Cardiac function indicators measured by echocardiography (LVEF (%), FS (%), LVPWd, IVSd, and LVIDd) in TAC- and sham-operated mice. (d) Expression of CTGF, periostin, and α -SMA mRNAs and proteins in heart cells ( n = 6). (e) Gene expression of P2X receptors in TAC- and sham-operated mice ( n = 6) at 4 weeks and gene expression of P2X7R in heart cells at 3 days, 1 week, 2 weeks, and 4 weeks after TAC- or sham-operated treatment ( n = 6). (f) Expression of P2X7R protein in mouse heart cells ( n = 6). Results are presented as means ± standard error of the mean. ∗ indicates P < 0.05, ∗∗ indicates P < 0.01, and ∗∗∗ indicates P < 0.001 versus sham.

    Article Snippet: The primary antibody for incubation was anti- α -SMA (Abcam, 1 : 200) or P2X7R (Alomone Labs, 1 : 200).

    Techniques: Staining, Expressing

    TGF- β 1-treatment upregulated P2X7R in CFs. (a) Immunofluorescence images showing α -SMA expression in CFs ( α -SMA (green) and nuclei DAPI (blue), n = 300). (b) Rate of cell proliferation (EdU-positive cells (red) and nuclei DAPI (blue), n = 120). (c) Expression of TGF- β , α -SMA, CTGF, and periostin mRNAs ( n = 3). (d) Expression of CTGF, α -SMA, periostin, and TGF- β proteins ( n = 3). (e) Western blot results showing P2X7R expression in CFs ( n = 3). (f) Immunofluorescence images showing P2RX7 expression in CFs (P2X7R (red) and nuclei DAPI (blue), n = 60). Results are presented as means ± standard error of the mean; ∗ indicates P < 0.05, ∗∗ indicates P < 0.01, and ∗∗∗ indicates P < 0.001 versus control.

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Inhibition of P2X7 Purinergic Receptor Ameliorates Cardiac Fibrosis by Suppressing NLRP3/IL-1 β Pathway

    doi: 10.1155/2020/7956274

    Figure Lengend Snippet: TGF- β 1-treatment upregulated P2X7R in CFs. (a) Immunofluorescence images showing α -SMA expression in CFs ( α -SMA (green) and nuclei DAPI (blue), n = 300). (b) Rate of cell proliferation (EdU-positive cells (red) and nuclei DAPI (blue), n = 120). (c) Expression of TGF- β , α -SMA, CTGF, and periostin mRNAs ( n = 3). (d) Expression of CTGF, α -SMA, periostin, and TGF- β proteins ( n = 3). (e) Western blot results showing P2X7R expression in CFs ( n = 3). (f) Immunofluorescence images showing P2RX7 expression in CFs (P2X7R (red) and nuclei DAPI (blue), n = 60). Results are presented as means ± standard error of the mean; ∗ indicates P < 0.05, ∗∗ indicates P < 0.01, and ∗∗∗ indicates P < 0.001 versus control.

    Article Snippet: The primary antibody for incubation was anti- α -SMA (Abcam, 1 : 200) or P2X7R (Alomone Labs, 1 : 200).

    Techniques: Immunofluorescence, Expressing, Western Blot

    Knockdown of P2X7R alleviates the effects of TGF- β 1 on CF activation. (a) Expression of P2X7R mRNA after siP2X7R transfection ( n = 6). (b) Expression of collagen I, CTGF, α -SMA, and TGF- β mRNAs after transfection of CFs with siP2X7R ( n = 6). (c, d) Expression of periostin, α -SMA, CTGF, P2X7R, NLRP3, IL-1 β , and caspase-1 proteins after transfection of CFs with siP2X7R ( n = 6). (e) Expression of α -SMA in CFs by immunofluorescence staining ( α -SMA (green) and nuclei DAPI (blue), n = 300). (f) Changes in CF proliferation (EdU-positive cells (red) and nuclei DAPI (blue), n = 120). (g) Expression of collagen I, CTGF, α -SMA, and TGF- β mRNAs after treatment of CFs with BBG ( n = 6). (h) Expression of P2X7R protein after BBG treatment ( n = 6). (i) Expression of periostin, α -SMA, CTGF, NLRP3, IL-1 β , and caspase-1 proteins after BBG treatment ( n = 6). (j) Expression of periostin, α -SMA, TGF- β and CTGF proteins in activated CFs after BzATP treatment ( n = 6). Results are presented as means ± standard error of the mean; ∗ indicates P < 0.05, ∗∗ indicates P < 0.01, and ∗∗∗ indicates P < 0.001 versus control or siNC; # indicates P < 0.05, ## indicates P < 0.01, and ### indicates P < 0.001.

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Inhibition of P2X7 Purinergic Receptor Ameliorates Cardiac Fibrosis by Suppressing NLRP3/IL-1 β Pathway

    doi: 10.1155/2020/7956274

    Figure Lengend Snippet: Knockdown of P2X7R alleviates the effects of TGF- β 1 on CF activation. (a) Expression of P2X7R mRNA after siP2X7R transfection ( n = 6). (b) Expression of collagen I, CTGF, α -SMA, and TGF- β mRNAs after transfection of CFs with siP2X7R ( n = 6). (c, d) Expression of periostin, α -SMA, CTGF, P2X7R, NLRP3, IL-1 β , and caspase-1 proteins after transfection of CFs with siP2X7R ( n = 6). (e) Expression of α -SMA in CFs by immunofluorescence staining ( α -SMA (green) and nuclei DAPI (blue), n = 300). (f) Changes in CF proliferation (EdU-positive cells (red) and nuclei DAPI (blue), n = 120). (g) Expression of collagen I, CTGF, α -SMA, and TGF- β mRNAs after treatment of CFs with BBG ( n = 6). (h) Expression of P2X7R protein after BBG treatment ( n = 6). (i) Expression of periostin, α -SMA, CTGF, NLRP3, IL-1 β , and caspase-1 proteins after BBG treatment ( n = 6). (j) Expression of periostin, α -SMA, TGF- β and CTGF proteins in activated CFs after BzATP treatment ( n = 6). Results are presented as means ± standard error of the mean; ∗ indicates P < 0.05, ∗∗ indicates P < 0.01, and ∗∗∗ indicates P < 0.001 versus control or siNC; # indicates P < 0.05, ## indicates P < 0.01, and ### indicates P < 0.001.

    Article Snippet: The primary antibody for incubation was anti- α -SMA (Abcam, 1 : 200) or P2X7R (Alomone Labs, 1 : 200).

    Techniques: Activation Assay, Expressing, Transfection, Immunofluorescence, Staining

    A schematic illustration of the mechanism of P2X7R in pressure overload-induced cardiac fibrosis and TGF- β 1-induced CF activation. In TAC-induced injury, pathological stress, such as pressure overload and TGF- β 1, triggers the release of ATP. Elevated ATP activates P2X7 receptors, contributing to P2X7-mediated NLRP3 inflammasome (NLRP3, ASC, and caspase-1) activation. This induces the release of IL-1 β , causing severe inflammation that precipitates cardiac fibrosis and aggravates TGF- β 1-induced CF activation. Moreover, inhibition of P2X7R with BBG inhibitor in TAC mice or CFs primed with TGF- β 1 reduces fibrotic extracellular matrix (ECM) gene expression and cardiac fibrosis.

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Inhibition of P2X7 Purinergic Receptor Ameliorates Cardiac Fibrosis by Suppressing NLRP3/IL-1 β Pathway

    doi: 10.1155/2020/7956274

    Figure Lengend Snippet: A schematic illustration of the mechanism of P2X7R in pressure overload-induced cardiac fibrosis and TGF- β 1-induced CF activation. In TAC-induced injury, pathological stress, such as pressure overload and TGF- β 1, triggers the release of ATP. Elevated ATP activates P2X7 receptors, contributing to P2X7-mediated NLRP3 inflammasome (NLRP3, ASC, and caspase-1) activation. This induces the release of IL-1 β , causing severe inflammation that precipitates cardiac fibrosis and aggravates TGF- β 1-induced CF activation. Moreover, inhibition of P2X7R with BBG inhibitor in TAC mice or CFs primed with TGF- β 1 reduces fibrotic extracellular matrix (ECM) gene expression and cardiac fibrosis.

    Article Snippet: The primary antibody for incubation was anti- α -SMA (Abcam, 1 : 200) or P2X7R (Alomone Labs, 1 : 200).

    Techniques: Activation Assay, Inhibition, Expressing

    Spleen cells from 3 to 4-mo-old MRL +/+ ( solid line ), MRL/ lpr ( dashed line ) and P2X7-deficient B6 P2X7R−/− ( dotted line ) mice (n = 3 mice/group) were treated for 45 min at 37°C with doses of ATP ranging from 100 to 5000 µM. Spleen cells were then triple-stained with anti-CD90, anti-CD19 and anti-CD62L mAb to assess by flow cytometry: (A) the percentage of CD62L-expressing CD19 – CD90 + T cells and (B) MFI of CD62L on CD19 – CD90 + T cells. Results are expressed as the mean percentage of initial expression ± SE.

    Journal: PLoS ONE

    Article Title: Loss of P2X7 Receptor Plasma Membrane Expression and Function in Pathogenic B220 + Double-Negative T Lymphocytes of Autoimmune MRL/ lpr Mice

    doi: 10.1371/journal.pone.0052161

    Figure Lengend Snippet: Spleen cells from 3 to 4-mo-old MRL +/+ ( solid line ), MRL/ lpr ( dashed line ) and P2X7-deficient B6 P2X7R−/− ( dotted line ) mice (n = 3 mice/group) were treated for 45 min at 37°C with doses of ATP ranging from 100 to 5000 µM. Spleen cells were then triple-stained with anti-CD90, anti-CD19 and anti-CD62L mAb to assess by flow cytometry: (A) the percentage of CD62L-expressing CD19 – CD90 + T cells and (B) MFI of CD62L on CD19 – CD90 + T cells. Results are expressed as the mean percentage of initial expression ± SE.

    Article Snippet: The membranes were blocked in TBS containing 3% nonfat dry milk and 0.2% Tween 20 for 1 h at room temperature and subsequently incubated overnight at 4°C with affinity purified rabbit anti-P2X7R antibodies recognizing residues 576 to 595 of P2X7R (1∶1000; Alomone Laboratories, Jerusalem, Israel).

    Techniques: Staining, Flow Cytometry, Expressing

    (A) The levels of CD39 expression on B220 – (either CD4 + or CD8 + ) (□) and B220 + DN (▪) T-cell subsets as well as on CD19 + B cells (▪) from MRL/ lpr mice (n = 3) were analyzed by flow cytometry. Results shown on histogram are normalized MFI (nMFI) calculated as MFI of positive cells × percentage of positive cells. Asterisks denote statistically significant differences between B220 + DN T cells or B220 – CD8 + T cells and B220 – CD4 + T cells: * p ≤0.05; ** p ≤0.01. (B) P2X7R mRNA expression was analyzed in FACS-purified B220 – (□) and B220 + (▪) CD19 – CD90 + T-cell subsets from individual B6, MRL +/+ and MRL/ lpr mice by real-time RT-PCR. Specific P2X7R cDNA was quantified by standard curves based on known amounts of PCR-amplified P2X7R cDNA. Data are expressed as amount (pg) of P2X7R RNA per cell, and histograms represent the mean ± SE of three independent experiments (n = 8 mice/group). (C) P2X7R protein levels in whole LN cells from 3 to 4-mo-old MRL +/+ , MRL/ lpr and B6 P2X7−/− mice, and FACS-sorted B220 – and B220 + CD19 – CD90 + T-cell subsets from MRL/ lpr mice were analyzed by western blotting using affinity-purified rabbit anti-P2X7R polyclonal antibodies (1∶1000; Alomone Laboratories). A non-specific band (*) is frequently observed with this polyclonal antibody . The blot was stripped and reprobed with anti-actin mAb. (D and E) Flow cytometric analysis of P2X7R expression levels on B220 – and B220 + T-cell subpopulations. Spleen cells from MRL +/+ , MRL/ lpr and B6 P2X7R−/− mice (n = 3 mice/group) were stained with anti-CD90 mAb, anti-B220 mAb, rabbit polyclonal anti-P2X7R antiserum (1∶100) generated by genetic immunization and with either anti-CD4 mAb, anti-CD8 mAb or anti-CD4 plus anti-CD8 mAbs. (D) Overlay histograms showing the expression levels of P2X7R on gated CD4 T cells or CD8 T cells from MRL +/+ mice (open histogram) and B6 P2X7R−/− mice (shaded histogram). (E) Overlay histograms comparing the expression levels of P2X7R on B220 – (–) and B220 + (–) T cells, either CD4 + ( left ) or CD8 + ( middle ), from MRL/ lpr mice. Right , overlay histogram comparing the expression levels of P2X7R on B220 + DN T cells (–) and B220 – CD4 + T cells (–) from MRL/ lpr mice. The results shown are representative of at least four independent experiments.

    Journal: PLoS ONE

    Article Title: Loss of P2X7 Receptor Plasma Membrane Expression and Function in Pathogenic B220 + Double-Negative T Lymphocytes of Autoimmune MRL/ lpr Mice

    doi: 10.1371/journal.pone.0052161

    Figure Lengend Snippet: (A) The levels of CD39 expression on B220 – (either CD4 + or CD8 + ) (□) and B220 + DN (▪) T-cell subsets as well as on CD19 + B cells (▪) from MRL/ lpr mice (n = 3) were analyzed by flow cytometry. Results shown on histogram are normalized MFI (nMFI) calculated as MFI of positive cells × percentage of positive cells. Asterisks denote statistically significant differences between B220 + DN T cells or B220 – CD8 + T cells and B220 – CD4 + T cells: * p ≤0.05; ** p ≤0.01. (B) P2X7R mRNA expression was analyzed in FACS-purified B220 – (□) and B220 + (▪) CD19 – CD90 + T-cell subsets from individual B6, MRL +/+ and MRL/ lpr mice by real-time RT-PCR. Specific P2X7R cDNA was quantified by standard curves based on known amounts of PCR-amplified P2X7R cDNA. Data are expressed as amount (pg) of P2X7R RNA per cell, and histograms represent the mean ± SE of three independent experiments (n = 8 mice/group). (C) P2X7R protein levels in whole LN cells from 3 to 4-mo-old MRL +/+ , MRL/ lpr and B6 P2X7−/− mice, and FACS-sorted B220 – and B220 + CD19 – CD90 + T-cell subsets from MRL/ lpr mice were analyzed by western blotting using affinity-purified rabbit anti-P2X7R polyclonal antibodies (1∶1000; Alomone Laboratories). A non-specific band (*) is frequently observed with this polyclonal antibody . The blot was stripped and reprobed with anti-actin mAb. (D and E) Flow cytometric analysis of P2X7R expression levels on B220 – and B220 + T-cell subpopulations. Spleen cells from MRL +/+ , MRL/ lpr and B6 P2X7R−/− mice (n = 3 mice/group) were stained with anti-CD90 mAb, anti-B220 mAb, rabbit polyclonal anti-P2X7R antiserum (1∶100) generated by genetic immunization and with either anti-CD4 mAb, anti-CD8 mAb or anti-CD4 plus anti-CD8 mAbs. (D) Overlay histograms showing the expression levels of P2X7R on gated CD4 T cells or CD8 T cells from MRL +/+ mice (open histogram) and B6 P2X7R−/− mice (shaded histogram). (E) Overlay histograms comparing the expression levels of P2X7R on B220 – (–) and B220 + (–) T cells, either CD4 + ( left ) or CD8 + ( middle ), from MRL/ lpr mice. Right , overlay histogram comparing the expression levels of P2X7R on B220 + DN T cells (–) and B220 – CD4 + T cells (–) from MRL/ lpr mice. The results shown are representative of at least four independent experiments.

    Article Snippet: The membranes were blocked in TBS containing 3% nonfat dry milk and 0.2% Tween 20 for 1 h at room temperature and subsequently incubated overnight at 4°C with affinity purified rabbit anti-P2X7R antibodies recognizing residues 576 to 595 of P2X7R (1∶1000; Alomone Laboratories, Jerusalem, Israel).

    Techniques: Expressing, Flow Cytometry, Purification, Quantitative RT-PCR, Amplification, Western Blot, Affinity Purification, Staining, Generated

    The sequences of oligonucleotide primers.

    Journal: BioMed Research International

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: The sequences of oligonucleotide primers.

    Article Snippet: Primary , Rabbit anti-P2X7R monoclonal IgG , 1 : 400 , Alomone Labs, Jerusalem, Israel.

    Techniques:

    The antibodies for immunoblotting.

    Journal: BioMed Research International

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: The antibodies for immunoblotting.

    Article Snippet: Primary , Rabbit anti-P2X7R monoclonal IgG , 1 : 400 , Alomone Labs, Jerusalem, Israel.

    Techniques: Western Blot, Concentration Assay

    The antibodies for fluorescence labeling.

    Journal: BioMed Research International

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: The antibodies for fluorescence labeling.

    Article Snippet: Primary , Rabbit anti-P2X7R monoclonal IgG , 1 : 400 , Alomone Labs, Jerusalem, Israel.

    Techniques: Fluorescence, Labeling, Concentration Assay

    Mechanical threshold and P2X7R and p-p38 expression. (a) Mechanical threshold after BTZ injection. (b, c) Western blot for P2X7R expression after BTZ treatment. (d) Immunofluorescence location of P2X7R in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. P2X7R is not expressed in NF-200-positive neurons. P2X7R is expressed in GFAP-labeled satellite glial cells (SGCs). (e) Immunofluorescence location of p-p38 in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. p-p38 is expressed in both MAP2-labeled neurons and GFAP-labeled SGCs. (f) Immunofluorescence location of P2X7R in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. P2X7R is expressed mainly in Iba-1-labeled microglial cells rather than in GFAP-labeled astrocytes and MAP2-labeled neurons. (g) Immunofluorescence location of p-p38 in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. p-p38 is expressed mainly in Iba-1-labeled microglial cells. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Journal: BioMed Research International

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: Mechanical threshold and P2X7R and p-p38 expression. (a) Mechanical threshold after BTZ injection. (b, c) Western blot for P2X7R expression after BTZ treatment. (d) Immunofluorescence location of P2X7R in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. P2X7R is not expressed in NF-200-positive neurons. P2X7R is expressed in GFAP-labeled satellite glial cells (SGCs). (e) Immunofluorescence location of p-p38 in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. p-p38 is expressed in both MAP2-labeled neurons and GFAP-labeled SGCs. (f) Immunofluorescence location of P2X7R in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. P2X7R is expressed mainly in Iba-1-labeled microglial cells rather than in GFAP-labeled astrocytes and MAP2-labeled neurons. (g) Immunofluorescence location of p-p38 in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. p-p38 is expressed mainly in Iba-1-labeled microglial cells. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Article Snippet: Primary , Rabbit anti-P2X7R monoclonal IgG , 1 : 400 , Alomone Labs, Jerusalem, Israel.

    Techniques: Expressing, Injection, Western Blot, Immunofluorescence, Labeling

    p38 mRNA expression and p38 phosphorylation in DRG after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) p-p38 immunofluorescence labeling. The arrows show the typical p-p38 single-labeled DRG cells. (e) p-p38 fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗∗ P < 0.001.

    Journal: BioMed Research International

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: p38 mRNA expression and p38 phosphorylation in DRG after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) p-p38 immunofluorescence labeling. The arrows show the typical p-p38 single-labeled DRG cells. (e) p-p38 fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗∗ P < 0.001.

    Article Snippet: Primary , Rabbit anti-P2X7R monoclonal IgG , 1 : 400 , Alomone Labs, Jerusalem, Israel.

    Techniques: Expressing, Inhibition, Western Blot, Immunofluorescence, Labeling, Fluorescence

    p38 mRNA expression and p38 phosphorylation in SDH after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Journal: BioMed Research International

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: p38 mRNA expression and p38 phosphorylation in SDH after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Article Snippet: Primary , Rabbit anti-P2X7R monoclonal IgG , 1 : 400 , Alomone Labs, Jerusalem, Israel.

    Techniques: Expressing, Inhibition, Western Blot, Fluorescence, Labeling

    IL-1 β , IL-6, and TNF- α mRNA expression in DRG and SDH after inhibition of P2X7R. (a) DRG IL-1 β mRNA. (b) DRG IL-6 mRNA. (c) DRG TNF- α mRNA. (d) SDH IL-1 β mRNA. (e) SDH IL-6 mRNA. (f) SDH TNF- α mRNA. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Journal: BioMed Research International

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: IL-1 β , IL-6, and TNF- α mRNA expression in DRG and SDH after inhibition of P2X7R. (a) DRG IL-1 β mRNA. (b) DRG IL-6 mRNA. (c) DRG TNF- α mRNA. (d) SDH IL-1 β mRNA. (e) SDH IL-6 mRNA. (f) SDH TNF- α mRNA. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Article Snippet: Primary , Rabbit anti-P2X7R monoclonal IgG , 1 : 400 , Alomone Labs, Jerusalem, Israel.

    Techniques: Expressing, Inhibition

    P2X7R mRNA and protein expression in DRG after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and GFAP coexpression fluorescence labeling for SGCs. The arrows indicate the typical single-labeled and double-labeled DRG satellite cells. (e) P2X7R and GFAP coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Journal: BioMed Research International

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: P2X7R mRNA and protein expression in DRG after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and GFAP coexpression fluorescence labeling for SGCs. The arrows indicate the typical single-labeled and double-labeled DRG satellite cells. (e) P2X7R and GFAP coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Article Snippet: Primary , Rabbit anti-P2X7R monoclonal IgG , 1 : 400 , Alomone Labs, Jerusalem, Israel.

    Techniques: Expressing, Inhibition, Western Blot, Fluorescence, Labeling

    P2X7R mRNA and protein expression in SDH after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 colocalization fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Journal: BioMed Research International

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: P2X7R mRNA and protein expression in SDH after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 colocalization fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Article Snippet: Primary , Rabbit anti-P2X7R monoclonal IgG , 1 : 400 , Alomone Labs, Jerusalem, Israel.

    Techniques: Expressing, Inhibition, Western Blot, Fluorescence, Labeling

    Mechanical threshold alterations after inhibition of P2X7R or p38. (a) Mechanical threshold after inhibition of P2X7R. (b) Mechanical threshold after inhibition of p38. Mean ± SEM ( n = 5). ∗∗∗ P < 0.001 (vs. control); # P < 0.05; ## P < 0.01 (vs. BTZ group).

    Journal: BioMed Research International

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: Mechanical threshold alterations after inhibition of P2X7R or p38. (a) Mechanical threshold after inhibition of P2X7R. (b) Mechanical threshold after inhibition of p38. Mean ± SEM ( n = 5). ∗∗∗ P < 0.001 (vs. control); # P < 0.05; ## P < 0.01 (vs. BTZ group).

    Article Snippet: Primary , Rabbit anti-P2X7R monoclonal IgG , 1 : 400 , Alomone Labs, Jerusalem, Israel.

    Techniques: Inhibition

    Immunofluorescence detected P2X7R of microglia in spinal cord. ( A ) P2X7R (P2X7R+, green) and IBA-1 (IBA-1+, red) co-expressed as P2X7R of microglia (P2X7R+/IBA-1+, yellow) that were overexpressed after spinal cord injury and BzATP intervention, and, compared to the morphology of microglia in the sham group, the morphology of activated microglia after injury changed to large soma and short irregular processes, revealing that many microglia in spinal cord became active after spinal cord injury. ( B ) The number of P2X7R of microglia (P2X7R+/IBA-1+) increased notably after spinal cord injury compared to the sham group (** P<0.01), and, compared to the model group, BzATP increased the number of P2X7R of microglia (* P<0.05) while A-438079 significantly decreased the number of P2X7R of microglia (** P<0.01). ( C ) The P2X7R-positive microglia percentage was clearly increased after spinal cord injury compared to the sham group (** P<0.01), and, compared to the model group, BzATP accounted for a significantly increased P2X7R-positive microglia proportion (** P<0.01) while A-438079 accounted for a significantly reduced P2X7R-positive microglia proportion (** P<0.01).

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    Article Title: P2X7 Receptor (P2X7R) of Microglia Mediates Neuroinflammation by Regulating (NOD)-Like Receptor Protein 3 (NLRP3) Inflammasome-Dependent Inflammation After Spinal Cord Injury

    doi: 10.12659/MSM.925491

    Figure Lengend Snippet: Immunofluorescence detected P2X7R of microglia in spinal cord. ( A ) P2X7R (P2X7R+, green) and IBA-1 (IBA-1+, red) co-expressed as P2X7R of microglia (P2X7R+/IBA-1+, yellow) that were overexpressed after spinal cord injury and BzATP intervention, and, compared to the morphology of microglia in the sham group, the morphology of activated microglia after injury changed to large soma and short irregular processes, revealing that many microglia in spinal cord became active after spinal cord injury. ( B ) The number of P2X7R of microglia (P2X7R+/IBA-1+) increased notably after spinal cord injury compared to the sham group (** P<0.01), and, compared to the model group, BzATP increased the number of P2X7R of microglia (* P<0.05) while A-438079 significantly decreased the number of P2X7R of microglia (** P<0.01). ( C ) The P2X7R-positive microglia percentage was clearly increased after spinal cord injury compared to the sham group (** P<0.01), and, compared to the model group, BzATP accounted for a significantly increased P2X7R-positive microglia proportion (** P<0.01) while A-438079 accounted for a significantly reduced P2X7R-positive microglia proportion (** P<0.01).

    Article Snippet: Sections were subjected to antigen retrieval with citric acid buffer, blocked with 10% normal goat serum for 2 h at room temperature, and incubated with a mixture of rabbit P2X7R antibody (1: 100, APR-004, Alomone, Jerusalem, Israel) and goat IBA-1 antibody (1: 200, ab5076, Abcam, Shanghai, China) at 4°C overnight.

    Techniques: Immunofluorescence

    Western blot and qPCR assay were used to assess the expression of P2X7R protein and P2X7R mRNA in spinal cord. ( A ) Western blot assay showed that the expression of P2X7R protein in spinal cord was increased notably after spinal cord injury compared to the sham group (** P<0.01) and, compared to model group, BzATP increased the expression of P2X7R protein in spinal cord (* P<0.05) while A-438079 significantly decreased the expression of P2X7R protein in spinal cord (** P<0.01). ( B ) qPCR assay showed that the expression of P2X7R mRNA in spinal cord increased notably after spinal cord injury compared to the sham group (** P<0.01), and compared to the model group, BzATP increased the expression of P2X7R mRNA in spinal cord (* P<0.05) while A-438079 significantly decreased the expression of P2X7R mRNA in spinal cord (** P<0.01).

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    Article Title: P2X7 Receptor (P2X7R) of Microglia Mediates Neuroinflammation by Regulating (NOD)-Like Receptor Protein 3 (NLRP3) Inflammasome-Dependent Inflammation After Spinal Cord Injury

    doi: 10.12659/MSM.925491

    Figure Lengend Snippet: Western blot and qPCR assay were used to assess the expression of P2X7R protein and P2X7R mRNA in spinal cord. ( A ) Western blot assay showed that the expression of P2X7R protein in spinal cord was increased notably after spinal cord injury compared to the sham group (** P<0.01) and, compared to model group, BzATP increased the expression of P2X7R protein in spinal cord (* P<0.05) while A-438079 significantly decreased the expression of P2X7R protein in spinal cord (** P<0.01). ( B ) qPCR assay showed that the expression of P2X7R mRNA in spinal cord increased notably after spinal cord injury compared to the sham group (** P<0.01), and compared to the model group, BzATP increased the expression of P2X7R mRNA in spinal cord (* P<0.05) while A-438079 significantly decreased the expression of P2X7R mRNA in spinal cord (** P<0.01).

    Article Snippet: Sections were subjected to antigen retrieval with citric acid buffer, blocked with 10% normal goat serum for 2 h at room temperature, and incubated with a mixture of rabbit P2X7R antibody (1: 100, APR-004, Alomone, Jerusalem, Israel) and goat IBA-1 antibody (1: 200, ab5076, Abcam, Shanghai, China) at 4°C overnight.

    Techniques: Western Blot, Expressing

    P2X7R of microglia in spinal cord-mediated neuroinflammation after spinal cord injury. ( A ) ELISA showed that, compared to the sham group, the expression of IL-1β and IL-18 in the spinal cord were significantly increased after spinal cord injury (** P<0.01) and, compared to the model group, A438079 significantly inhibited the expression of IL- 1β and IL-18 (** P<0.01) and BzATP significantly increased the expression of IL- 1β and IL-18 (** P<0.01), which was strongly inhibited by CY-09 (** P<0.01). ( B ) qPCR assay showed that, compared to the sham group, the expression of IL-1β mRNA and IL-18 mRNA in the spinal cord was significantly increased after spinal cord injury (** P<0.01) and, compared to the model group, A438079 significantly inhibited the expression of IL-1β mRNA and IL-18 mRNA (** P<0.01), and BzATP significantly increased the expression of IL-1β mRNA and IL-18 mRNA (** P<0.01), which was strongly inhibited by CY-09 (** P<0.01).

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    Article Title: P2X7 Receptor (P2X7R) of Microglia Mediates Neuroinflammation by Regulating (NOD)-Like Receptor Protein 3 (NLRP3) Inflammasome-Dependent Inflammation After Spinal Cord Injury

    doi: 10.12659/MSM.925491

    Figure Lengend Snippet: P2X7R of microglia in spinal cord-mediated neuroinflammation after spinal cord injury. ( A ) ELISA showed that, compared to the sham group, the expression of IL-1β and IL-18 in the spinal cord were significantly increased after spinal cord injury (** P<0.01) and, compared to the model group, A438079 significantly inhibited the expression of IL- 1β and IL-18 (** P<0.01) and BzATP significantly increased the expression of IL- 1β and IL-18 (** P<0.01), which was strongly inhibited by CY-09 (** P<0.01). ( B ) qPCR assay showed that, compared to the sham group, the expression of IL-1β mRNA and IL-18 mRNA in the spinal cord was significantly increased after spinal cord injury (** P<0.01) and, compared to the model group, A438079 significantly inhibited the expression of IL-1β mRNA and IL-18 mRNA (** P<0.01), and BzATP significantly increased the expression of IL-1β mRNA and IL-18 mRNA (** P<0.01), which was strongly inhibited by CY-09 (** P<0.01).

    Article Snippet: Sections were subjected to antigen retrieval with citric acid buffer, blocked with 10% normal goat serum for 2 h at room temperature, and incubated with a mixture of rabbit P2X7R antibody (1: 100, APR-004, Alomone, Jerusalem, Israel) and goat IBA-1 antibody (1: 200, ab5076, Abcam, Shanghai, China) at 4°C overnight.

    Techniques: Enzyme-linked Immunosorbent Assay, Expressing

    P2X7R of microglia in spinal cord regulated the expression of NLRP3 and cleaved-Caspase-1 (P20). Compared to the sham group, the expression of NLRP3 and cleaved-Caspase-1 (P20) in the spinal cord significantly increased after spinal cord injury (** P<0.01) and, compared to the model group, BzATP promoted the expression of NLRP3 and cleaved-Caspase-1 (P20) (* P<0.05), which was inhibited by CY-09 (* P<0.05), while A-438079 notably reduced the expression of NLRP3 and cleaved-Caspase-1 (P20) (** P<0.01).

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    Article Title: P2X7 Receptor (P2X7R) of Microglia Mediates Neuroinflammation by Regulating (NOD)-Like Receptor Protein 3 (NLRP3) Inflammasome-Dependent Inflammation After Spinal Cord Injury

    doi: 10.12659/MSM.925491

    Figure Lengend Snippet: P2X7R of microglia in spinal cord regulated the expression of NLRP3 and cleaved-Caspase-1 (P20). Compared to the sham group, the expression of NLRP3 and cleaved-Caspase-1 (P20) in the spinal cord significantly increased after spinal cord injury (** P<0.01) and, compared to the model group, BzATP promoted the expression of NLRP3 and cleaved-Caspase-1 (P20) (* P<0.05), which was inhibited by CY-09 (* P<0.05), while A-438079 notably reduced the expression of NLRP3 and cleaved-Caspase-1 (P20) (** P<0.01).

    Article Snippet: Sections were subjected to antigen retrieval with citric acid buffer, blocked with 10% normal goat serum for 2 h at room temperature, and incubated with a mixture of rabbit P2X7R antibody (1: 100, APR-004, Alomone, Jerusalem, Israel) and goat IBA-1 antibody (1: 200, ab5076, Abcam, Shanghai, China) at 4°C overnight.

    Techniques: Expressing