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  • 93
    Alomone Labs anti-p2x7 receptor (extracellular)-fitc antibody
    Anti P2x7 Receptor (Extracellular) Fitc Antibody, 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/anti-p2x7 receptor (extracellular)-fitc antibody/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    93
    Alomone Labs anti p2x7r extracellular fitc
    P24XR and <t>P2X7R</t> Mediate ATP-Dependent Calcium Signal Propagation (A and B) Murine RAW 264.7 macrophages were loaded with photoactivable caged-IP 3 and the fluorescent calcium indicator Fluo-4 (green), and calcium signal propagation after IP 3 uncaging in the origin cell (white box) was monitored in live imaging. Experiments were performed in HBSS with 2 mM Ca 2+ (A) or in calcium-free HBSS supplemented with 2 mM EGTA (B). Scale bars, 50 μm. See also . (C and D) Quantification of 3 independent live calcium imaging experiments with RAW 264.7 (C) or primary BMDMs (D). Error bars represent SEM. For statistical data analysis, Student’s t test was used ( ∗ p < 0.05). (E and F) Relative mRNA expression of different members of the P2X family of receptors, measured by real-time PCR in the RAW 264.7 cell line (E) and in BMDM primary macrophages (F). Error bars represent SEM. (G) Quantification of 5 independent live calcium imaging experiments with RAW 264.7 cells pre-treated with the P2X4R inhibitor 5BDBD (100 μM, 30 min at 37°C), the P2X7R inhibitor A740003 (100 μM, 30 min at 37°C), or their vehicle (DMSO) or left untreated. Error bars represent SEM. For statistical data analysis, One-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant). (H) Representative western blot (left) and quantification of repeated experiments (right) of P2X4R and P2X7R expression in RAW 264.7 cells transfected with siRNA specific for P2X4R and P2X7R or with scramble siRNA. Control cells were electroporated in the absence of oligonucleotides. (I) Quantification of 3 independent live calcium imaging experiments with RAW 264.7 cells silenced for P2X4R and P2X7R. Error bars represent SEM. For statistical data analysis, one-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant).
    Anti P2x7r Extracellular Fitc, 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/anti p2x7r extracellular fitc/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti p2x7r extracellular fitc - by Bioz Stars, 2024-07
    93/100 stars
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    93
    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, 2024-07
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    93
    Alomone Labs p2x 7 catalogue number apr 008 f fluorescein isothiocyanate
    Model of pore former induced lysis. A bacterial toxin inserts a large channel or pore into the erythrocyte membrane. ATP is immediately released through the pore and activates <t>P2X</t> receptors. The membrane insertion of the toxin also causes a steep rise in the intracellular Ca 2+ concentration ([Ca 2+ ] i ), which results from Ca 2+ passing through the pore itself and from activation of P2X receptors, which are non-selective cation channels permeable to Ca 2+ . The increase in [Ca 2+ ] i activates the Ca 2+ -sensitive K + channel K Ca 3.1 and Cl − channel TMEM16A, which results in K + and Cl − efflux and cell shrinkage as obligated water follows. The cells will remain shrunken as long as the K + efflux surpasses the Na + influx via the toxin pore and the P2X receptors. Prolonged stimulation of <t>P2X</t> <t>7</t> can activate pannexins, which will contribute to the Na + influx. Eventually, the Na + influx will exceed the K + efflux and the cells will swell and eventually burst. Blockage of the P2X 1 and P2X 7 receptor has been proven as a protective measure for bacterial toxins, and for complement to carry out their toxicity. The model based on previous work [ , , , ]
    P2x 7 Catalogue Number Apr 008 F Fluorescein Isothiocyanate, 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/p2x 7 catalogue number apr 008 f fluorescein isothiocyanate/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p2x 7 catalogue number apr 008 f fluorescein isothiocyanate - by Bioz Stars, 2024-07
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    93
    Alomone Labs extracellular atp
    Model of pore former induced lysis. A bacterial toxin inserts a large channel or pore into the erythrocyte membrane. ATP is immediately released through the pore and activates <t>P2X</t> receptors. The membrane insertion of the toxin also causes a steep rise in the intracellular Ca 2+ concentration ([Ca 2+ ] i ), which results from Ca 2+ passing through the pore itself and from activation of P2X receptors, which are non-selective cation channels permeable to Ca 2+ . The increase in [Ca 2+ ] i activates the Ca 2+ -sensitive K + channel K Ca 3.1 and Cl − channel TMEM16A, which results in K + and Cl − efflux and cell shrinkage as obligated water follows. The cells will remain shrunken as long as the K + efflux surpasses the Na + influx via the toxin pore and the P2X receptors. Prolonged stimulation of <t>P2X</t> <t>7</t> can activate pannexins, which will contribute to the Na + influx. Eventually, the Na + influx will exceed the K + efflux and the cells will swell and eventually burst. Blockage of the P2X 1 and P2X 7 receptor has been proven as a protective measure for bacterial toxins, and for complement to carry out their toxicity. The model based on previous work [ , , , ]
    Extracellular Atp, 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/extracellular atp/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    extracellular atp - by Bioz Stars, 2024-07
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    Image Search Results


    P24XR and P2X7R Mediate ATP-Dependent Calcium Signal Propagation (A and B) Murine RAW 264.7 macrophages were loaded with photoactivable caged-IP 3 and the fluorescent calcium indicator Fluo-4 (green), and calcium signal propagation after IP 3 uncaging in the origin cell (white box) was monitored in live imaging. Experiments were performed in HBSS with 2 mM Ca 2+ (A) or in calcium-free HBSS supplemented with 2 mM EGTA (B). Scale bars, 50 μm. See also . (C and D) Quantification of 3 independent live calcium imaging experiments with RAW 264.7 (C) or primary BMDMs (D). Error bars represent SEM. For statistical data analysis, Student’s t test was used ( ∗ p < 0.05). (E and F) Relative mRNA expression of different members of the P2X family of receptors, measured by real-time PCR in the RAW 264.7 cell line (E) and in BMDM primary macrophages (F). Error bars represent SEM. (G) Quantification of 5 independent live calcium imaging experiments with RAW 264.7 cells pre-treated with the P2X4R inhibitor 5BDBD (100 μM, 30 min at 37°C), the P2X7R inhibitor A740003 (100 μM, 30 min at 37°C), or their vehicle (DMSO) or left untreated. Error bars represent SEM. For statistical data analysis, One-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant). (H) Representative western blot (left) and quantification of repeated experiments (right) of P2X4R and P2X7R expression in RAW 264.7 cells transfected with siRNA specific for P2X4R and P2X7R or with scramble siRNA. Control cells were electroporated in the absence of oligonucleotides. (I) Quantification of 3 independent live calcium imaging experiments with RAW 264.7 cells silenced for P2X4R and P2X7R. Error bars represent SEM. For statistical data analysis, one-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant).

    Journal: Cell Reports

    Article Title: Intercellular Calcium Signaling Induced by ATP Potentiates Macrophage Phagocytosis

    doi: 10.1016/j.celrep.2019.03.011

    Figure Lengend Snippet: P24XR and P2X7R Mediate ATP-Dependent Calcium Signal Propagation (A and B) Murine RAW 264.7 macrophages were loaded with photoactivable caged-IP 3 and the fluorescent calcium indicator Fluo-4 (green), and calcium signal propagation after IP 3 uncaging in the origin cell (white box) was monitored in live imaging. Experiments were performed in HBSS with 2 mM Ca 2+ (A) or in calcium-free HBSS supplemented with 2 mM EGTA (B). Scale bars, 50 μm. See also . (C and D) Quantification of 3 independent live calcium imaging experiments with RAW 264.7 (C) or primary BMDMs (D). Error bars represent SEM. For statistical data analysis, Student’s t test was used ( ∗ p < 0.05). (E and F) Relative mRNA expression of different members of the P2X family of receptors, measured by real-time PCR in the RAW 264.7 cell line (E) and in BMDM primary macrophages (F). Error bars represent SEM. (G) Quantification of 5 independent live calcium imaging experiments with RAW 264.7 cells pre-treated with the P2X4R inhibitor 5BDBD (100 μM, 30 min at 37°C), the P2X7R inhibitor A740003 (100 μM, 30 min at 37°C), or their vehicle (DMSO) or left untreated. Error bars represent SEM. For statistical data analysis, One-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant). (H) Representative western blot (left) and quantification of repeated experiments (right) of P2X4R and P2X7R expression in RAW 264.7 cells transfected with siRNA specific for P2X4R and P2X7R or with scramble siRNA. Control cells were electroporated in the absence of oligonucleotides. (I) Quantification of 3 independent live calcium imaging experiments with RAW 264.7 cells silenced for P2X4R and P2X7R. Error bars represent SEM. For statistical data analysis, one-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant).

    Article Snippet: After counting, cells were incubated with anti-CD16/CD32 (BD PharMingen), and subsequently stained with the appropriate combinations of the following antibodies: anti-Cd11b-PerCP Cy5.5 (M1/70, BD Biosciences), anti-CD169-AlexaFluor647 (MOMA-1, Biorad), anti-P2X7R-extracellular-FITC (Alomone Labs), purified anti-P2X4R-extracellular (Alomone Labs) followed by incubation with the secondary anti-rabbit-FITC antibody (Thermo Fisher).

    Techniques: Imaging, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Transfection

    Macrophage Polarization Status Affects Calcium Signal Propagation (A) The surface expression of P2X4R (top) and P2X7R (bottom) was analyzed by flow cytometry in resting, IFNγ-treated (10 ng/mL, 24 h), or IL4-treated (20 ng/mL, 24 h) macrophages. Histograms show the quantification 3 independent biological replicates. Error bars represent SEM. For data analysis, one-way ANOVA followed by Bonferroni’s multiple comparisons test was used (ns, non-significant; ∗∗ p < 0.01; ∗∗∗ p < 0.001). (B) Maximal back-projection of two representative live calcium imaging experiments performed with IFNγ- or IL4-treated RAW 264.7 cells loaded with caged-IP 3 and Fluo-4. The fluorescence variation 60 s after the irradiation of the origin cell (white box) is represented in false colors. (C) Representative traces of live calcium imaging experiments, showing the fluorescence variation after the uncaging of the origin cell (red) and the bystander macrophages (black). (D) Quantification of 3 independent live calcium imaging experiments. Error bars represent SEM. For data analysis, one-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant).

    Journal: Cell Reports

    Article Title: Intercellular Calcium Signaling Induced by ATP Potentiates Macrophage Phagocytosis

    doi: 10.1016/j.celrep.2019.03.011

    Figure Lengend Snippet: Macrophage Polarization Status Affects Calcium Signal Propagation (A) The surface expression of P2X4R (top) and P2X7R (bottom) was analyzed by flow cytometry in resting, IFNγ-treated (10 ng/mL, 24 h), or IL4-treated (20 ng/mL, 24 h) macrophages. Histograms show the quantification 3 independent biological replicates. Error bars represent SEM. For data analysis, one-way ANOVA followed by Bonferroni’s multiple comparisons test was used (ns, non-significant; ∗∗ p < 0.01; ∗∗∗ p < 0.001). (B) Maximal back-projection of two representative live calcium imaging experiments performed with IFNγ- or IL4-treated RAW 264.7 cells loaded with caged-IP 3 and Fluo-4. The fluorescence variation 60 s after the irradiation of the origin cell (white box) is represented in false colors. (C) Representative traces of live calcium imaging experiments, showing the fluorescence variation after the uncaging of the origin cell (red) and the bystander macrophages (black). (D) Quantification of 3 independent live calcium imaging experiments. Error bars represent SEM. For data analysis, one-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant).

    Article Snippet: After counting, cells were incubated with anti-CD16/CD32 (BD PharMingen), and subsequently stained with the appropriate combinations of the following antibodies: anti-Cd11b-PerCP Cy5.5 (M1/70, BD Biosciences), anti-CD169-AlexaFluor647 (MOMA-1, Biorad), anti-P2X7R-extracellular-FITC (Alomone Labs), purified anti-P2X4R-extracellular (Alomone Labs) followed by incubation with the secondary anti-rabbit-FITC antibody (Thermo Fisher).

    Techniques: Expressing, Flow Cytometry, Imaging, Fluorescence, Irradiation

    Extracellular ATP Is Required for Efficient Phagocytosis (A) Primary BMDMs were incubated with PhRodo E. coli fluorescent bioparticles in the presence or absence of 5 mM EGTA to chelate extracellular calcium. Phagocytosis was monitored at 15 or 30 min by flow cytometry (see <xref ref-type=Figure S4 ). Macrophages incubated with 20 μM cytochalasin D were used as negative reference. The phagocytic index was calculated as the percentage of fluorescent macrophages multiplied by their mean of fluorescence (MFI) and normalized on the cytochalasin-treated samples. (B) Primary BMDMs were loaded with the intracellular calcium chelator BAPTA-AM or its vehicle (loading solution) before performing the phagocytosis assay. (C) Primary BMDMs were incubated with PhRodo E. coli , PhRodo Zymosan, or PhRodo S. aureus fluorescent bioparticles, in the presence or absence of apyrase (5 U/mL). (D) Primary BMDMs were pretreated with the P2X4R inhibitor 5BDBD (100 μM), the P2X7R inhibitor A740003 (100 μM), or their vehicle (DMSO), or were left untreated, before performing the phagocytosis assay. (E) Phagocytosis was performed for 30 min in the presence or absence of MSC-derived EVs, pre-incubated or not with ARL-67516 (30 min, 200 μM). The graphs are representative of at least 3 independent biological replicates, each performed in technical triplicate. Error bars represent SEM. For data analysis, a two-way ANOVA followed by Tukey’s multiple comparisons test was used ( ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ns, non-significant). " width="100%" height="100%">

    Journal: Cell Reports

    Article Title: Intercellular Calcium Signaling Induced by ATP Potentiates Macrophage Phagocytosis

    doi: 10.1016/j.celrep.2019.03.011

    Figure Lengend Snippet: Extracellular ATP Is Required for Efficient Phagocytosis (A) Primary BMDMs were incubated with PhRodo E. coli fluorescent bioparticles in the presence or absence of 5 mM EGTA to chelate extracellular calcium. Phagocytosis was monitored at 15 or 30 min by flow cytometry (see Figure S4 ). Macrophages incubated with 20 μM cytochalasin D were used as negative reference. The phagocytic index was calculated as the percentage of fluorescent macrophages multiplied by their mean of fluorescence (MFI) and normalized on the cytochalasin-treated samples. (B) Primary BMDMs were loaded with the intracellular calcium chelator BAPTA-AM or its vehicle (loading solution) before performing the phagocytosis assay. (C) Primary BMDMs were incubated with PhRodo E. coli , PhRodo Zymosan, or PhRodo S. aureus fluorescent bioparticles, in the presence or absence of apyrase (5 U/mL). (D) Primary BMDMs were pretreated with the P2X4R inhibitor 5BDBD (100 μM), the P2X7R inhibitor A740003 (100 μM), or their vehicle (DMSO), or were left untreated, before performing the phagocytosis assay. (E) Phagocytosis was performed for 30 min in the presence or absence of MSC-derived EVs, pre-incubated or not with ARL-67516 (30 min, 200 μM). The graphs are representative of at least 3 independent biological replicates, each performed in technical triplicate. Error bars represent SEM. For data analysis, a two-way ANOVA followed by Tukey’s multiple comparisons test was used ( ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ns, non-significant).

    Article Snippet: After counting, cells were incubated with anti-CD16/CD32 (BD PharMingen), and subsequently stained with the appropriate combinations of the following antibodies: anti-Cd11b-PerCP Cy5.5 (M1/70, BD Biosciences), anti-CD169-AlexaFluor647 (MOMA-1, Biorad), anti-P2X7R-extracellular-FITC (Alomone Labs), purified anti-P2X4R-extracellular (Alomone Labs) followed by incubation with the secondary anti-rabbit-FITC antibody (Thermo Fisher).

    Techniques: Incubation, Flow Cytometry, Fluorescence, Phagocytosis Assay, Derivative Assay

    Journal: Cell Reports

    Article Title: Intercellular Calcium Signaling Induced by ATP Potentiates Macrophage Phagocytosis

    doi: 10.1016/j.celrep.2019.03.011

    Figure Lengend Snippet:

    Article Snippet: After counting, cells were incubated with anti-CD16/CD32 (BD PharMingen), and subsequently stained with the appropriate combinations of the following antibodies: anti-Cd11b-PerCP Cy5.5 (M1/70, BD Biosciences), anti-CD169-AlexaFluor647 (MOMA-1, Biorad), anti-P2X7R-extracellular-FITC (Alomone Labs), purified anti-P2X4R-extracellular (Alomone Labs) followed by incubation with the secondary anti-rabbit-FITC antibody (Thermo Fisher).

    Techniques: Recombinant, Negative Control, Real-time Polymerase Chain Reaction, Software

    Journal: Cell Reports

    Article Title: Intercellular Calcium Signaling Induced by ATP Potentiates Macrophage Phagocytosis

    doi: 10.1016/j.celrep.2019.03.011

    Figure Lengend Snippet:

    Article Snippet: After counting, cells were incubated with anti-CD16/CD32 (BD PharMingen), and subsequently stained with the appropriate combinations of the following antibodies: anti-Cd11b-PerCP Cy5.5 (M1/70, BD Biosciences), anti-CD169-AlexaFluor647 (MOMA-1, Biorad), anti-P2X7R-extracellular-FITC (Alomone Labs), purified anti-P2X4R-extracellular (Alomone Labs) followed by incubation with the secondary anti-rabbit-FITC antibody (Thermo Fisher).

    Techniques:

    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

    Model of pore former induced lysis. A bacterial toxin inserts a large channel or pore into the erythrocyte membrane. ATP is immediately released through the pore and activates P2X receptors. The membrane insertion of the toxin also causes a steep rise in the intracellular Ca 2+ concentration ([Ca 2+ ] i ), which results from Ca 2+ passing through the pore itself and from activation of P2X receptors, which are non-selective cation channels permeable to Ca 2+ . The increase in [Ca 2+ ] i activates the Ca 2+ -sensitive K + channel K Ca 3.1 and Cl − channel TMEM16A, which results in K + and Cl − efflux and cell shrinkage as obligated water follows. The cells will remain shrunken as long as the K + efflux surpasses the Na + influx via the toxin pore and the P2X receptors. Prolonged stimulation of P2X 7 can activate pannexins, which will contribute to the Na + influx. Eventually, the Na + influx will exceed the K + efflux and the cells will swell and eventually burst. Blockage of the P2X 1 and P2X 7 receptor has been proven as a protective measure for bacterial toxins, and for complement to carry out their toxicity. The model based on previous work [ , , , ]

    Journal: Critical Care

    Article Title: Erythrocyte P2X 1 receptor expression is correlated with change in haematocrit in patients admitted to the ICU with blood pathogen-positive sepsis

    doi: 10.1186/s13054-018-2100-3

    Figure Lengend Snippet: Model of pore former induced lysis. A bacterial toxin inserts a large channel or pore into the erythrocyte membrane. ATP is immediately released through the pore and activates P2X receptors. The membrane insertion of the toxin also causes a steep rise in the intracellular Ca 2+ concentration ([Ca 2+ ] i ), which results from Ca 2+ passing through the pore itself and from activation of P2X receptors, which are non-selective cation channels permeable to Ca 2+ . The increase in [Ca 2+ ] i activates the Ca 2+ -sensitive K + channel K Ca 3.1 and Cl − channel TMEM16A, which results in K + and Cl − efflux and cell shrinkage as obligated water follows. The cells will remain shrunken as long as the K + efflux surpasses the Na + influx via the toxin pore and the P2X receptors. Prolonged stimulation of P2X 7 can activate pannexins, which will contribute to the Na + influx. Eventually, the Na + influx will exceed the K + efflux and the cells will swell and eventually burst. Blockage of the P2X 1 and P2X 7 receptor has been proven as a protective measure for bacterial toxins, and for complement to carry out their toxicity. The model based on previous work [ , , , ]

    Article Snippet: Cells in each sample were counted on a cell counter (Spectre, Merch Millipore, USA) prior to exposure to either P2X 1 (catalogue number APR-022-AG) or P2X 7 (catalogue number APR-008-F) fluorescein isothiocyanate (FITC)-conjugated antibody (Alomone Labs, Jerusalem, Israel) for 1 h at room temperature in the dark at 200 rpm in concentrations according to the manufacturer’s instructions (10 mg antibody to 10 6 cells).

    Techniques: Lysis, Concentration Assay, Activation Assay

    Freeze-thaw - control experiments. a Whole blood drawn from healthy volunteers was incubated either with vehicle (Veh), 100 μM NF449 (P2X 1 antagonist) or 100 μM A804598 (P2X 7 antagonist) and frozen and stored for 3 weeks as whole blood at − 80 °C. After thawing, lysis was measured as absorbance of free haemoglobin in supernatant at 540 nm. n = 3, mean ± SEM. b Flow cytometry of thawed whole blood samples diluted 1000-fold, to ascertain equal distribution of blood cell sub-populations. Cells were identified by forward scatter FSC and side scatter SSC and quantified based on fixed regions. c Bar graph shows data presented as mean ± SEM, n = 3. ns, not significant

    Journal: Critical Care

    Article Title: Erythrocyte P2X 1 receptor expression is correlated with change in haematocrit in patients admitted to the ICU with blood pathogen-positive sepsis

    doi: 10.1186/s13054-018-2100-3

    Figure Lengend Snippet: Freeze-thaw - control experiments. a Whole blood drawn from healthy volunteers was incubated either with vehicle (Veh), 100 μM NF449 (P2X 1 antagonist) or 100 μM A804598 (P2X 7 antagonist) and frozen and stored for 3 weeks as whole blood at − 80 °C. After thawing, lysis was measured as absorbance of free haemoglobin in supernatant at 540 nm. n = 3, mean ± SEM. b Flow cytometry of thawed whole blood samples diluted 1000-fold, to ascertain equal distribution of blood cell sub-populations. Cells were identified by forward scatter FSC and side scatter SSC and quantified based on fixed regions. c Bar graph shows data presented as mean ± SEM, n = 3. ns, not significant

    Article Snippet: Cells in each sample were counted on a cell counter (Spectre, Merch Millipore, USA) prior to exposure to either P2X 1 (catalogue number APR-022-AG) or P2X 7 (catalogue number APR-008-F) fluorescein isothiocyanate (FITC)-conjugated antibody (Alomone Labs, Jerusalem, Israel) for 1 h at room temperature in the dark at 200 rpm in concentrations according to the manufacturer’s instructions (10 mg antibody to 10 6 cells).

    Techniques: Incubation, Lysis, Flow Cytometry

    Correlation statistics

    Journal: Critical Care

    Article Title: Erythrocyte P2X 1 receptor expression is correlated with change in haematocrit in patients admitted to the ICU with blood pathogen-positive sepsis

    doi: 10.1186/s13054-018-2100-3

    Figure Lengend Snippet: Correlation statistics

    Article Snippet: Cells in each sample were counted on a cell counter (Spectre, Merch Millipore, USA) prior to exposure to either P2X 1 (catalogue number APR-022-AG) or P2X 7 (catalogue number APR-008-F) fluorescein isothiocyanate (FITC)-conjugated antibody (Alomone Labs, Jerusalem, Israel) for 1 h at room temperature in the dark at 200 rpm in concentrations according to the manufacturer’s instructions (10 mg antibody to 10 6 cells).

    Techniques: Activity Assay, Expressing

    Haematocrit and haemoglobin levels and erythrocyte P2X 7 receptor expression. a Change in haematocrit in blood pathogen-positive patients with sepsis between Emergency Department (ER) and ICU admission and P2X 7 receptor expression on erythrocytes were not correlated. The right panel shows the change in haematocrit in blood pathogen-positive patients grouped by high or low expression of P2X 7 (Δhct/hour). b Change in haematocrit in blood pathogen-negative patients with sepsis between ER and ICU admission and P2X 7 receptor expression on erythrocytes. Right panel shows the change in haematocrit in blood pathogen-negative patients grouped by high or low expression of P2X 7 (Δhct/hour). c Change in haemoglobin in blood pathogen-positive patients with sepsis between ER and ICU admission and P2X 7 receptor expression on erythrocytes were not correlated. The right panel shows change in haemoglobin in blood pathogen-positive patients grouped by high or low expression of P2X 7 (ΔHgb/hour, not significant (ns)). d Change in haemoglobin in blood pathogen-negative patients with sepsis between ER and ICU admission and P2X 7 receptor expression on erythrocytes. The right panel shows change in haemoglobin in blood pathogen-negative patients grouped by high or low expression of P2X 7 (ΔHgb/hour)

    Journal: Critical Care

    Article Title: Erythrocyte P2X 1 receptor expression is correlated with change in haematocrit in patients admitted to the ICU with blood pathogen-positive sepsis

    doi: 10.1186/s13054-018-2100-3

    Figure Lengend Snippet: Haematocrit and haemoglobin levels and erythrocyte P2X 7 receptor expression. a Change in haematocrit in blood pathogen-positive patients with sepsis between Emergency Department (ER) and ICU admission and P2X 7 receptor expression on erythrocytes were not correlated. The right panel shows the change in haematocrit in blood pathogen-positive patients grouped by high or low expression of P2X 7 (Δhct/hour). b Change in haematocrit in blood pathogen-negative patients with sepsis between ER and ICU admission and P2X 7 receptor expression on erythrocytes. Right panel shows the change in haematocrit in blood pathogen-negative patients grouped by high or low expression of P2X 7 (Δhct/hour). c Change in haemoglobin in blood pathogen-positive patients with sepsis between ER and ICU admission and P2X 7 receptor expression on erythrocytes were not correlated. The right panel shows change in haemoglobin in blood pathogen-positive patients grouped by high or low expression of P2X 7 (ΔHgb/hour, not significant (ns)). d Change in haemoglobin in blood pathogen-negative patients with sepsis between ER and ICU admission and P2X 7 receptor expression on erythrocytes. The right panel shows change in haemoglobin in blood pathogen-negative patients grouped by high or low expression of P2X 7 (ΔHgb/hour)

    Article Snippet: Cells in each sample were counted on a cell counter (Spectre, Merch Millipore, USA) prior to exposure to either P2X 1 (catalogue number APR-022-AG) or P2X 7 (catalogue number APR-008-F) fluorescein isothiocyanate (FITC)-conjugated antibody (Alomone Labs, Jerusalem, Israel) for 1 h at room temperature in the dark at 200 rpm in concentrations according to the manufacturer’s instructions (10 mg antibody to 10 6 cells).

    Techniques: Expressing