p2x7  (Alomone Labs)


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    Structured Review

    Alomone Labs p2x7
    <t>P2X7</t> receptor promotes invadopodial activity of extracellular matrix degradation. ( a ) 4T1 mouse mammary and MDA-MB-435s human cancer cells were grown on Matrigel™ containing DQ-BSA as a fluorogenic substrate for proteases, emitting red fluorescence when degraded. P2X7 receptor was immunodetected using a rabbit primary antibody against P2X7 protein and a secondary ant-rabbit IgG antibody conjugated to AF488. The right panel shows the fluorescence values for the two channels at the locations indicated by a blue arrow. Scale bar, 100 µm. ( b ) Invadopodia entrapped into a 2%-gelatin matrix were fractionated and separated from cytosol and membranes-enriched fractions. The quality of the fractions was assessed by western blotting after SDS-PAGE, using cytosolic (HSC70), membrane (caveolin-1, β-adaptin) and invadopodia (cathepsin B, cortactin, focal adhesion kinase) markers. P2X7 proteins were found to be present in the membrane fractions and enriched in the invadopodia fraction. This figure is representative of 5 independent experiments. The full image blots can be found in Figure S5 . ( c ) The invadopodial activity was assessed as being F-actin foci (green labelling, phalloidin-488) co-localised with focused proteolytic activities (red labelling, DQ-BSA proteolysis) from 4T1 cells grown for 24 h on Matrigel™ containing DQ-BSA in control condition (vehicle), or stimulated with 300 µM BzATP, in the presence and absence of 10 µM A438079. “Coloc” indicates co-localization of degradative activity and F-actin foci and appears as white pixels. Scale bar, 50 µm. ( d ) The number of degradation areas per cell was counted in 20 images, from five independent experiments. * indicates a statistically significant difference at p
    P2x7, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p2x7/product/Alomone Labs
    Average 93 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    p2x7 - by Bioz Stars, 2022-12
    93/100 stars

    Images

    1) Product Images from "P2X7 Receptor Promotes Mouse Mammary Cancer Cell Invasiveness and Tumour Progression, and Is a Target for Anticancer Treatment"

    Article Title: P2X7 Receptor Promotes Mouse Mammary Cancer Cell Invasiveness and Tumour Progression, and Is a Target for Anticancer Treatment

    Journal: Cancers

    doi: 10.3390/cancers12092342

    P2X7 receptor promotes invadopodial activity of extracellular matrix degradation. ( a ) 4T1 mouse mammary and MDA-MB-435s human cancer cells were grown on Matrigel™ containing DQ-BSA as a fluorogenic substrate for proteases, emitting red fluorescence when degraded. P2X7 receptor was immunodetected using a rabbit primary antibody against P2X7 protein and a secondary ant-rabbit IgG antibody conjugated to AF488. The right panel shows the fluorescence values for the two channels at the locations indicated by a blue arrow. Scale bar, 100 µm. ( b ) Invadopodia entrapped into a 2%-gelatin matrix were fractionated and separated from cytosol and membranes-enriched fractions. The quality of the fractions was assessed by western blotting after SDS-PAGE, using cytosolic (HSC70), membrane (caveolin-1, β-adaptin) and invadopodia (cathepsin B, cortactin, focal adhesion kinase) markers. P2X7 proteins were found to be present in the membrane fractions and enriched in the invadopodia fraction. This figure is representative of 5 independent experiments. The full image blots can be found in Figure S5 . ( c ) The invadopodial activity was assessed as being F-actin foci (green labelling, phalloidin-488) co-localised with focused proteolytic activities (red labelling, DQ-BSA proteolysis) from 4T1 cells grown for 24 h on Matrigel™ containing DQ-BSA in control condition (vehicle), or stimulated with 300 µM BzATP, in the presence and absence of 10 µM A438079. “Coloc” indicates co-localization of degradative activity and F-actin foci and appears as white pixels. Scale bar, 50 µm. ( d ) The number of degradation areas per cell was counted in 20 images, from five independent experiments. * indicates a statistically significant difference at p
    Figure Legend Snippet: P2X7 receptor promotes invadopodial activity of extracellular matrix degradation. ( a ) 4T1 mouse mammary and MDA-MB-435s human cancer cells were grown on Matrigel™ containing DQ-BSA as a fluorogenic substrate for proteases, emitting red fluorescence when degraded. P2X7 receptor was immunodetected using a rabbit primary antibody against P2X7 protein and a secondary ant-rabbit IgG antibody conjugated to AF488. The right panel shows the fluorescence values for the two channels at the locations indicated by a blue arrow. Scale bar, 100 µm. ( b ) Invadopodia entrapped into a 2%-gelatin matrix were fractionated and separated from cytosol and membranes-enriched fractions. The quality of the fractions was assessed by western blotting after SDS-PAGE, using cytosolic (HSC70), membrane (caveolin-1, β-adaptin) and invadopodia (cathepsin B, cortactin, focal adhesion kinase) markers. P2X7 proteins were found to be present in the membrane fractions and enriched in the invadopodia fraction. This figure is representative of 5 independent experiments. The full image blots can be found in Figure S5 . ( c ) The invadopodial activity was assessed as being F-actin foci (green labelling, phalloidin-488) co-localised with focused proteolytic activities (red labelling, DQ-BSA proteolysis) from 4T1 cells grown for 24 h on Matrigel™ containing DQ-BSA in control condition (vehicle), or stimulated with 300 µM BzATP, in the presence and absence of 10 µM A438079. “Coloc” indicates co-localization of degradative activity and F-actin foci and appears as white pixels. Scale bar, 50 µm. ( d ) The number of degradation areas per cell was counted in 20 images, from five independent experiments. * indicates a statistically significant difference at p

    Techniques Used: Activity Assay, Multiple Displacement Amplification, Fluorescence, Western Blot, SDS Page

    P2X7 antagonism slows mammary tumour growth. ( a ) In vivo mouse experiments assessing the effects of treatments with two P2X7 antagonist (A438079 or AZ10606120) on primary tumour growth, in which cancer cells and host organisms both express the P2rx7 gene. ( b ) primary mammary tumour growth represented as a function of the duration of the treatment depending on antagonist treatments (A438079 or AZ10606120) compared to the injection of vehicle. ( c – e ) A Gompertz model was used to assess mammary tumour growth as a function of time for ( c ), vehicle group, ( d ), A438079 and ( e ), AZ10606120. Black, red and blue dots represent individual tumour volumes of these respective categories, blue areas are 90% model prediction intervals and lines are the median tumour growth. Estimated mean (inter-individual standard deviation) of model parameters were k growth = 0.64 day-1 (39%), V max = 1.620 mm 3 (64%), EFF = 0.52 (82%) and γ = 0.17 (–). This analysis showed that tumour volume doubled in a mean time of 0.64 day, that the mean maximum tumour volume was 1.620 mm 3 and that tumour growth was twice slower in the presence of A438079 or AZ10606120 treatment ( p
    Figure Legend Snippet: P2X7 antagonism slows mammary tumour growth. ( a ) In vivo mouse experiments assessing the effects of treatments with two P2X7 antagonist (A438079 or AZ10606120) on primary tumour growth, in which cancer cells and host organisms both express the P2rx7 gene. ( b ) primary mammary tumour growth represented as a function of the duration of the treatment depending on antagonist treatments (A438079 or AZ10606120) compared to the injection of vehicle. ( c – e ) A Gompertz model was used to assess mammary tumour growth as a function of time for ( c ), vehicle group, ( d ), A438079 and ( e ), AZ10606120. Black, red and blue dots represent individual tumour volumes of these respective categories, blue areas are 90% model prediction intervals and lines are the median tumour growth. Estimated mean (inter-individual standard deviation) of model parameters were k growth = 0.64 day-1 (39%), V max = 1.620 mm 3 (64%), EFF = 0.52 (82%) and γ = 0.17 (–). This analysis showed that tumour volume doubled in a mean time of 0.64 day, that the mean maximum tumour volume was 1.620 mm 3 and that tumour growth was twice slower in the presence of A438079 or AZ10606120 treatment ( p

    Techniques Used: In Vivo, Injection, Standard Deviation

    P2X7 receptor in mammary cancer cells enhances primary tumour growth and metastatic development in vivo. ( a ) P2rx7 mRNA expression levels assessed by RT-qPCR in CTL, Crispr#1 and Crispr#2 cell lines, expressed relatively to 4T1 cells. * indicated a statistically significant difference from CTL cells at p
    Figure Legend Snippet: P2X7 receptor in mammary cancer cells enhances primary tumour growth and metastatic development in vivo. ( a ) P2rx7 mRNA expression levels assessed by RT-qPCR in CTL, Crispr#1 and Crispr#2 cell lines, expressed relatively to 4T1 cells. * indicated a statistically significant difference from CTL cells at p

    Techniques Used: In Vivo, Expressing, Quantitative RT-PCR, CRISPR

    P2X7 receptor stimulation promotes the acquisition of a mesenchymal phenotype. ( a ) F-actin cytoskeleton was visualized using phalloidin-488 in 4T1 cells stimulated with 300 µM BzATP at the time indicated. Rapid morphological changes appear as rapidly as after 6 min stimulation. Scale bar, 25 µm. ( b ) The number of new filopodia per cell was counted per hour in cells transfected with the LifeAct plasmid and studied in the presence of 300 µM BzATP, in the presence or absence of 10 µM A438079. * indicates a statistically significant difference at p
    Figure Legend Snippet: P2X7 receptor stimulation promotes the acquisition of a mesenchymal phenotype. ( a ) F-actin cytoskeleton was visualized using phalloidin-488 in 4T1 cells stimulated with 300 µM BzATP at the time indicated. Rapid morphological changes appear as rapidly as after 6 min stimulation. Scale bar, 25 µm. ( b ) The number of new filopodia per cell was counted per hour in cells transfected with the LifeAct plasmid and studied in the presence of 300 µM BzATP, in the presence or absence of 10 µM A438079. * indicates a statistically significant difference at p

    Techniques Used: Transfection, Plasmid Preparation

    2) Product Images from "Atheroprone flow activates inflammation via endothelial ATP-dependent P2X7-p38 signalling"

    Article Title: Atheroprone flow activates inflammation via endothelial ATP-dependent P2X7-p38 signalling

    Journal: Cardiovascular Research

    doi: 10.1093/cvr/cvx213

    P2X7 receptor activity occurs exclusively in HUVEC under atheroprone flow. Representative single cell ATP-induced calcium responses in atheroprotected ( A ) or atheroprone ( B, C ) flow conditioned HUVEC after thapsigargin pre-treatment ± the P2X7 inhibitor 10 µM A438079 (P2X7i) or siRNA control and P2X7 knockdown ( C ) and analysed by measuring the average area under the curve ( n = 4, 175 cells per donor, * indicates P ≤ 0.05 using a paired t -test). Values are mean ± S.E.M.
    Figure Legend Snippet: P2X7 receptor activity occurs exclusively in HUVEC under atheroprone flow. Representative single cell ATP-induced calcium responses in atheroprotected ( A ) or atheroprone ( B, C ) flow conditioned HUVEC after thapsigargin pre-treatment ± the P2X7 inhibitor 10 µM A438079 (P2X7i) or siRNA control and P2X7 knockdown ( C ) and analysed by measuring the average area under the curve ( n = 4, 175 cells per donor, * indicates P ≤ 0.05 using a paired t -test). Values are mean ± S.E.M.

    Techniques Used: Activity Assay

    Expression of ATP-gated P2X7 receptors is enhanced under atheroprone flow. ( A ) Western blot and densitometry of P2X7 in flow conditioned HUVEC using the ibidi flow system ( n = 6, ** indicates P ≤ 0.01 using a paired t -test). P2X7 is shown by an arrow, validated by P2X7 knockdown, shown in Supplementary material online , Figure 4 B. ( B ) Western blot and densitometry of P2X7 in flow conditioned HUVEC using the orbital shaker system ( n = 4, ** indicates P ≤ 0.01 using a paired t -test). ( C ) Representative en face immunostaining for P2X7 (red) on wildtype C57BL/6 mice at atheroprotective (outer curvature) and atheroprone (inner curvature) regions of the aorta (scale = 25 µm). Endothelial cells were identified by staining with CD31 (green) and cell nuclei were stained using TO-PRO (blue). P2X7 expression was analysed by measuring the relative fluorescent intensity at sites protected or prone to atherosclerosis. Surface expression was measured by measuring P2X7 fluorescence at sites co-localized with the endothelial cell surface marker CD31. Relative fluorescent intensities for P2X7 were corrected against the relative fluorescent intensity of IgG performed on the descending aorta ( n = 5, * indicates P ≤ 0.05 and *** indicates P ≤ 0.001 using a paired t -test). Values are mean ± S.E.M.
    Figure Legend Snippet: Expression of ATP-gated P2X7 receptors is enhanced under atheroprone flow. ( A ) Western blot and densitometry of P2X7 in flow conditioned HUVEC using the ibidi flow system ( n = 6, ** indicates P ≤ 0.01 using a paired t -test). P2X7 is shown by an arrow, validated by P2X7 knockdown, shown in Supplementary material online , Figure 4 B. ( B ) Western blot and densitometry of P2X7 in flow conditioned HUVEC using the orbital shaker system ( n = 4, ** indicates P ≤ 0.01 using a paired t -test). ( C ) Representative en face immunostaining for P2X7 (red) on wildtype C57BL/6 mice at atheroprotective (outer curvature) and atheroprone (inner curvature) regions of the aorta (scale = 25 µm). Endothelial cells were identified by staining with CD31 (green) and cell nuclei were stained using TO-PRO (blue). P2X7 expression was analysed by measuring the relative fluorescent intensity at sites protected or prone to atherosclerosis. Surface expression was measured by measuring P2X7 fluorescence at sites co-localized with the endothelial cell surface marker CD31. Relative fluorescent intensities for P2X7 were corrected against the relative fluorescent intensity of IgG performed on the descending aorta ( n = 5, * indicates P ≤ 0.05 and *** indicates P ≤ 0.001 using a paired t -test). Values are mean ± S.E.M.

    Techniques Used: Expressing, Western Blot, Immunostaining, Mouse Assay, Staining, Fluorescence, Marker

    P2X7 regulates E-selectin expression at sites susceptible to atherosclerosis in vivo . Representative en face immunostaining for E-selectin (red) on wildtype or P2X7 −/− BALB/c mice at atheroprotective (outer curvature) and atheroprone (inner curvature) sites of the aorta ( n = 4–5) (scale = 25 µm). Endothelial cells were identified by staining with CD31 (green) and cell nuclei were stained using TO-PRO (blue). E-selectin expression was analysed by measuring the relative fluorescent intensity at sites of protected or prone to atherosclerosis ( n = 4–5, * indicates P ≤ 0.05, ** indicates P ≤ 0.01 and *** indicates P ≤ 0.001 using a two-way ANOVA). Relative fluorescent intensities for P2X7 were corrected against the relative fluorescent intensity of IgG. Values are mean ± S.E.M.
    Figure Legend Snippet: P2X7 regulates E-selectin expression at sites susceptible to atherosclerosis in vivo . Representative en face immunostaining for E-selectin (red) on wildtype or P2X7 −/− BALB/c mice at atheroprotective (outer curvature) and atheroprone (inner curvature) sites of the aorta ( n = 4–5) (scale = 25 µm). Endothelial cells were identified by staining with CD31 (green) and cell nuclei were stained using TO-PRO (blue). E-selectin expression was analysed by measuring the relative fluorescent intensity at sites of protected or prone to atherosclerosis ( n = 4–5, * indicates P ≤ 0.05, ** indicates P ≤ 0.01 and *** indicates P ≤ 0.001 using a two-way ANOVA). Relative fluorescent intensities for P2X7 were corrected against the relative fluorescent intensity of IgG. Values are mean ± S.E.M.

    Techniques Used: Expressing, In Vivo, Immunostaining, Mouse Assay, Staining

    Atheroprone flow mediated inflammatory signalling is regulated by P2X7 responses. qPCR analysis of E-selectin ( A ) and IL-8 ( B ) in HUVEC conditioned with atheroprotective or atheroprone flow using the ibidi system ± the P2X7 inhibitor 10 µM A438079 (P2X7i) ( n = 5, * indicates P ≤ 0.05 using a two-way ANOVA). ( C ) IL-8 release measured by ELISA in the supernatant of HUVEC conditioned under atheroprotective or atheroprone flow using the ibidi system ± the P2X7 inhibitor 10 µM A438079 (P2X7i) ( n = 9, * indicates P ≤ 0.05, *** indicates p =
    Figure Legend Snippet: Atheroprone flow mediated inflammatory signalling is regulated by P2X7 responses. qPCR analysis of E-selectin ( A ) and IL-8 ( B ) in HUVEC conditioned with atheroprotective or atheroprone flow using the ibidi system ± the P2X7 inhibitor 10 µM A438079 (P2X7i) ( n = 5, * indicates P ≤ 0.05 using a two-way ANOVA). ( C ) IL-8 release measured by ELISA in the supernatant of HUVEC conditioned under atheroprotective or atheroprone flow using the ibidi system ± the P2X7 inhibitor 10 µM A438079 (P2X7i) ( n = 9, * indicates P ≤ 0.05, *** indicates p =

    Techniques Used: Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

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    Alomone Labs anti p2x7 receptor extracellular antibody
    QSG inhibited the <t>P2X7R-NEK7-NLRP3</t> inflammasome pathway. ( A ) Determination of potassium ion concentration in RAW264.7 macrophages supernatant. ( B ) The effective concentration range of CCK8 to detect AZ. ( C ) Representative graph of optimal concentration of AZ for WB validation. ( D ) Representative immunofluorescence detection images of RAW264.7 macrophages P2X7R expression. Scale bar=25 µm. ( E – H ) Western blot analysis showed that both QSG and AZ treatment reduced the expression of P2X7R, Caspase-1, IL-18, IL-1β in RAW264.7 macrophages, N = 3 per group. ### P
    Anti P2x7 Receptor Extracellular Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p2x7 receptor extracellular antibody/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti p2x7 receptor extracellular antibody - by Bioz Stars, 2022-12
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    Alomone Labs anti p2x7 receptor extracellular fitc antibody
    Tregs and <t>anti-inflammatory</t> cytokines in <t>P2X7R-regulated</t> acute gouty arthritis. ( A ) The expressions of Tregs among the three groups at 12h. ( B ) The levels of CD4 + CD25 + FOXP3 + Tregs in three groups at each time point. ( C) The expression of FOXP3 mRNA among three groups at each time point. ( D -E) The expressions of TGF-β 1 and IL-10 in serum among the three groups. *P
    Anti P2x7 Receptor Extracellular Fitc Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p2x7 receptor extracellular fitc antibody/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti p2x7 receptor extracellular fitc antibody - by Bioz Stars, 2022-12
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    Image Search Results


    QSG inhibited the P2X7R-NEK7-NLRP3 inflammasome pathway. ( A ) Determination of potassium ion concentration in RAW264.7 macrophages supernatant. ( B ) The effective concentration range of CCK8 to detect AZ. ( C ) Representative graph of optimal concentration of AZ for WB validation. ( D ) Representative immunofluorescence detection images of RAW264.7 macrophages P2X7R expression. Scale bar=25 µm. ( E – H ) Western blot analysis showed that both QSG and AZ treatment reduced the expression of P2X7R, Caspase-1, IL-18, IL-1β in RAW264.7 macrophages, N = 3 per group. ### P

    Journal: Journal of Inflammation Research

    Article Title: P2X7R-NEK7-NLRP3 Inflammasome Activation: A Novel Therapeutic Pathway of Qishen Granule in the Treatment of Acute Myocardial Ischemia

    doi: 10.2147/JIR.S373962

    Figure Lengend Snippet: QSG inhibited the P2X7R-NEK7-NLRP3 inflammasome pathway. ( A ) Determination of potassium ion concentration in RAW264.7 macrophages supernatant. ( B ) The effective concentration range of CCK8 to detect AZ. ( C ) Representative graph of optimal concentration of AZ for WB validation. ( D ) Representative immunofluorescence detection images of RAW264.7 macrophages P2X7R expression. Scale bar=25 µm. ( E – H ) Western blot analysis showed that both QSG and AZ treatment reduced the expression of P2X7R, Caspase-1, IL-18, IL-1β in RAW264.7 macrophages, N = 3 per group. ### P

    Article Snippet: The primary antibodies used are as follows: anti-Caspase-1 antibody (ab1872, Abcam, USA), anti-P2X7R antibody (APR-008, Alomone, Israel).

    Techniques: Concentration Assay, Western Blot, Immunofluorescence, Expressing

    QSG inhibited LPS combined with ATP-induced NLRP3 inflammasome activation in RAW264.7 macrophages. ( A ) Establishment of LPS combined with ATP to induce NLRP3 inflammasome activation model. ( B ) Immunofluorescence determined that LPS in combination with ATP activates NLRP3 inflammasome. Scale bar=75 µm. ( C ) Safe concentration of QSG in RAW264.7 macrophages. ( D ) Experimental protocol for QSG in LPS combined with ATP to induce NLRP3 inflammasome activation in RAW264.7 macrophages. ( E ) Effective concentration of QSG in RAW264.7 macrophages. ( F ) MCC950 inhibitor concentration range in RAW264.7 macrophages. ( G – M ) Western blot analysis showed that QSG treatment reduced the expression of P2X7R, NEK7, NLRP3, ASC, Caspase-1, IL-18, IL-1β in RAW264.7 macrophages. N = 3 per group. ## P

    Journal: Journal of Inflammation Research

    Article Title: P2X7R-NEK7-NLRP3 Inflammasome Activation: A Novel Therapeutic Pathway of Qishen Granule in the Treatment of Acute Myocardial Ischemia

    doi: 10.2147/JIR.S373962

    Figure Lengend Snippet: QSG inhibited LPS combined with ATP-induced NLRP3 inflammasome activation in RAW264.7 macrophages. ( A ) Establishment of LPS combined with ATP to induce NLRP3 inflammasome activation model. ( B ) Immunofluorescence determined that LPS in combination with ATP activates NLRP3 inflammasome. Scale bar=75 µm. ( C ) Safe concentration of QSG in RAW264.7 macrophages. ( D ) Experimental protocol for QSG in LPS combined with ATP to induce NLRP3 inflammasome activation in RAW264.7 macrophages. ( E ) Effective concentration of QSG in RAW264.7 macrophages. ( F ) MCC950 inhibitor concentration range in RAW264.7 macrophages. ( G – M ) Western blot analysis showed that QSG treatment reduced the expression of P2X7R, NEK7, NLRP3, ASC, Caspase-1, IL-18, IL-1β in RAW264.7 macrophages. N = 3 per group. ## P

    Article Snippet: The primary antibodies used are as follows: anti-Caspase-1 antibody (ab1872, Abcam, USA), anti-P2X7R antibody (APR-008, Alomone, Israel).

    Techniques: Activation Assay, Immunofluorescence, Concentration Assay, Western Blot, Expressing

    QSG inhibited NLRP3 inflamat some activation in cardiac tissue macrophages. ( A ) Representative NLRP3/ASC/Caspase-1 immunofluorescent staining images of AMI mice in each group. Scale bar=20 µm. ( B ) Representative CD86/Caspase-1 immunofluorescent staining images of AMI mice in each group. Scale bar=20 µm. ( C – I ) Western blot analysis showed that QSG treatment reduced the expression of P2X7R, NEK7, NLRP3, ASC, Caspase-1, IL-18, IL-1β in cardiac tissue macrophages. Proteins had been normalized to GAPDH, N = 6 per group. ### P

    Journal: Journal of Inflammation Research

    Article Title: P2X7R-NEK7-NLRP3 Inflammasome Activation: A Novel Therapeutic Pathway of Qishen Granule in the Treatment of Acute Myocardial Ischemia

    doi: 10.2147/JIR.S373962

    Figure Lengend Snippet: QSG inhibited NLRP3 inflamat some activation in cardiac tissue macrophages. ( A ) Representative NLRP3/ASC/Caspase-1 immunofluorescent staining images of AMI mice in each group. Scale bar=20 µm. ( B ) Representative CD86/Caspase-1 immunofluorescent staining images of AMI mice in each group. Scale bar=20 µm. ( C – I ) Western blot analysis showed that QSG treatment reduced the expression of P2X7R, NEK7, NLRP3, ASC, Caspase-1, IL-18, IL-1β in cardiac tissue macrophages. Proteins had been normalized to GAPDH, N = 6 per group. ### P

    Article Snippet: The primary antibodies used are as follows: anti-Caspase-1 antibody (ab1872, Abcam, USA), anti-P2X7R antibody (APR-008, Alomone, Israel).

    Techniques: Activation Assay, Staining, Mouse Assay, Western Blot, Expressing

    QSG might protect AMI through the P2X7R-NEK7-NLRP3 pathway.

    Journal: Journal of Inflammation Research

    Article Title: P2X7R-NEK7-NLRP3 Inflammasome Activation: A Novel Therapeutic Pathway of Qishen Granule in the Treatment of Acute Myocardial Ischemia

    doi: 10.2147/JIR.S373962

    Figure Lengend Snippet: QSG might protect AMI through the P2X7R-NEK7-NLRP3 pathway.

    Article Snippet: The primary antibodies used are as follows: anti-Caspase-1 antibody (ab1872, Abcam, USA), anti-P2X7R antibody (APR-008, Alomone, Israel).

    Techniques:

    QSG regulates the P2X7R-NEK7-NLRP3 pathway. ( A ) mP2X7R pcDNA3.1–3xFlag-T2A-EGFP map. ( B ) WB images of images of RAW264.7 macrophages and analysis of P2X7R. ( C ) Representative immunofluorescence detection images of Caspase-1. Scale bar=75 µm. ( D and E ) Western blot images of RAW264.7 macrophages and analysis of IL-18 and IL-1β. ( F – H ) RAW264.7 macrophages were transfected with P2X7R pcDNA3.1 followed by immunoprecipitation and immunoblot using NEK7 antibody to analyze the protein expression levels of NEK7, NLRP3. The Input represents total protein extracts used for immunoprecipitation. GAPDH was used as a loading control. IB, immunoblot. IP, immunoprecipitation. IgG, negative control. ### P

    Journal: Journal of Inflammation Research

    Article Title: P2X7R-NEK7-NLRP3 Inflammasome Activation: A Novel Therapeutic Pathway of Qishen Granule in the Treatment of Acute Myocardial Ischemia

    doi: 10.2147/JIR.S373962

    Figure Lengend Snippet: QSG regulates the P2X7R-NEK7-NLRP3 pathway. ( A ) mP2X7R pcDNA3.1–3xFlag-T2A-EGFP map. ( B ) WB images of images of RAW264.7 macrophages and analysis of P2X7R. ( C ) Representative immunofluorescence detection images of Caspase-1. Scale bar=75 µm. ( D and E ) Western blot images of RAW264.7 macrophages and analysis of IL-18 and IL-1β. ( F – H ) RAW264.7 macrophages were transfected with P2X7R pcDNA3.1 followed by immunoprecipitation and immunoblot using NEK7 antibody to analyze the protein expression levels of NEK7, NLRP3. The Input represents total protein extracts used for immunoprecipitation. GAPDH was used as a loading control. IB, immunoblot. IP, immunoprecipitation. IgG, negative control. ### P

    Article Snippet: The primary antibodies used are as follows: anti-Caspase-1 antibody (ab1872, Abcam, USA), anti-P2X7R antibody (APR-008, Alomone, Israel).

    Techniques: Western Blot, Immunofluorescence, Transfection, Immunoprecipitation, Expressing, Negative Control

    Tregs and anti-inflammatory cytokines in P2X7R-regulated acute gouty arthritis. ( A ) The expressions of Tregs among the three groups at 12h. ( B ) The levels of CD4 + CD25 + FOXP3 + Tregs in three groups at each time point. ( C) The expression of FOXP3 mRNA among three groups at each time point. ( D -E) The expressions of TGF-β 1 and IL-10 in serum among the three groups. *P

    Journal: Journal of Inflammation Research

    Article Title: ATP-Activated P2X7R Promote the Attack of Acute Gouty Arthritis in Rats Through Activating NLRP3 Inflammasome and Inflammatory Cytokine Production

    doi: 10.2147/JIR.S351660

    Figure Lengend Snippet: Tregs and anti-inflammatory cytokines in P2X7R-regulated acute gouty arthritis. ( A ) The expressions of Tregs among the three groups at 12h. ( B ) The levels of CD4 + CD25 + FOXP3 + Tregs in three groups at each time point. ( C) The expression of FOXP3 mRNA among three groups at each time point. ( D -E) The expressions of TGF-β 1 and IL-10 in serum among the three groups. *P

    Article Snippet: The treated spleen cells (1×10^6cells) were stained with anti-P2X7 receptor (extracellular)-FITC antibody (Alomone, Jerusalem, Israel) and CD68 antibody (NOVUS, Littleton, USA), and the white cell suspension obtained from the spleen was stained with FITC conjugated mouse anti-rat CD4, PE conjugated mouse anti-rat CD25 (BD Bioscience, MD, USA), PE conjugated anti-IL-17A and APC conjugated anti-Foxp3 antibodies (eBioscience, San Jose, CA, USA).

    Techniques: Expressing

    P2X7R regulates the development of acute gouty arthritis in rats. ( A ) At 12h, clinical manifestations of right ankle joint in rats among ATP group, BBG group and control group. ( B and C ) Clinical score and swelling index of three groups of rats at each time point. ( D ) Inflammatory cells infiltrated the synovial tissue of the right ankle joint in three groups of rats, at 24h. ( E and F ) Infiltration of mononuclear cells and neutrophils in the synovial tissue of the ankle joint among the three groups. * P

    Journal: Journal of Inflammation Research

    Article Title: ATP-Activated P2X7R Promote the Attack of Acute Gouty Arthritis in Rats Through Activating NLRP3 Inflammasome and Inflammatory Cytokine Production

    doi: 10.2147/JIR.S351660

    Figure Lengend Snippet: P2X7R regulates the development of acute gouty arthritis in rats. ( A ) At 12h, clinical manifestations of right ankle joint in rats among ATP group, BBG group and control group. ( B and C ) Clinical score and swelling index of three groups of rats at each time point. ( D ) Inflammatory cells infiltrated the synovial tissue of the right ankle joint in three groups of rats, at 24h. ( E and F ) Infiltration of mononuclear cells and neutrophils in the synovial tissue of the ankle joint among the three groups. * P

    Article Snippet: The treated spleen cells (1×10^6cells) were stained with anti-P2X7 receptor (extracellular)-FITC antibody (Alomone, Jerusalem, Israel) and CD68 antibody (NOVUS, Littleton, USA), and the white cell suspension obtained from the spleen was stained with FITC conjugated mouse anti-rat CD4, PE conjugated mouse anti-rat CD25 (BD Bioscience, MD, USA), PE conjugated anti-IL-17A and APC conjugated anti-Foxp3 antibodies (eBioscience, San Jose, CA, USA).

    Techniques:

    P2X7R promote Th17 cells and pro-inflammatory cytokines production. ( A ) The expressions of CD4 + IL-17 + Th17 cells among the three groups at 12h. ( B ) The levels of CD4 + IL-17 + Th17 cells in ATP group, BBG group and Control group at each time point. ( C-E ) The serum levels of IL-1β, IL-6 and IL-17 of the three groups at each time point. (F) The expression of IL-17 mRNA among three groups at each time point. * P

    Journal: Journal of Inflammation Research

    Article Title: ATP-Activated P2X7R Promote the Attack of Acute Gouty Arthritis in Rats Through Activating NLRP3 Inflammasome and Inflammatory Cytokine Production

    doi: 10.2147/JIR.S351660

    Figure Lengend Snippet: P2X7R promote Th17 cells and pro-inflammatory cytokines production. ( A ) The expressions of CD4 + IL-17 + Th17 cells among the three groups at 12h. ( B ) The levels of CD4 + IL-17 + Th17 cells in ATP group, BBG group and Control group at each time point. ( C-E ) The serum levels of IL-1β, IL-6 and IL-17 of the three groups at each time point. (F) The expression of IL-17 mRNA among three groups at each time point. * P

    Article Snippet: The treated spleen cells (1×10^6cells) were stained with anti-P2X7 receptor (extracellular)-FITC antibody (Alomone, Jerusalem, Israel) and CD68 antibody (NOVUS, Littleton, USA), and the white cell suspension obtained from the spleen was stained with FITC conjugated mouse anti-rat CD4, PE conjugated mouse anti-rat CD25 (BD Bioscience, MD, USA), PE conjugated anti-IL-17A and APC conjugated anti-Foxp3 antibodies (eBioscience, San Jose, CA, USA).

    Techniques: Expressing

    P2X7R mediated NLRP3 inflammatory-dependent IL-1β secretion on macrophages. The expressions of P2X7R ( A ), NLRP3 ( B ) and IL-1β ( C ) in rat spleen macrophages were analyzed by qRT-PCR. *P

    Journal: Journal of Inflammation Research

    Article Title: ATP-Activated P2X7R Promote the Attack of Acute Gouty Arthritis in Rats Through Activating NLRP3 Inflammasome and Inflammatory Cytokine Production

    doi: 10.2147/JIR.S351660

    Figure Lengend Snippet: P2X7R mediated NLRP3 inflammatory-dependent IL-1β secretion on macrophages. The expressions of P2X7R ( A ), NLRP3 ( B ) and IL-1β ( C ) in rat spleen macrophages were analyzed by qRT-PCR. *P

    Article Snippet: The treated spleen cells (1×10^6cells) were stained with anti-P2X7 receptor (extracellular)-FITC antibody (Alomone, Jerusalem, Israel) and CD68 antibody (NOVUS, Littleton, USA), and the white cell suspension obtained from the spleen was stained with FITC conjugated mouse anti-rat CD4, PE conjugated mouse anti-rat CD25 (BD Bioscience, MD, USA), PE conjugated anti-IL-17A and APC conjugated anti-Foxp3 antibodies (eBioscience, San Jose, CA, USA).

    Techniques: Quantitative RT-PCR

    The expression changes of P2X7R on macrophages affect their ability to take up YO-PRO-1. ( A ) At 12h, the expression level of P2X7R in each group of macrophages was detected by flow cytometry. ( B ) Expression of P2X7R among the three groups at different time points. ( C ) At 12h, the percentage of YO-PRO-1 uptake by macrophages in each group was detected by flow cytometry. ( D ) The ability of macrophages to uptake YO-PRO-1 among the three groups at each time point. ( E ) The expression level of P2X7R was positively correlated with the ability of macrophages to uptake YO-PRO-1. * P

    Journal: Journal of Inflammation Research

    Article Title: ATP-Activated P2X7R Promote the Attack of Acute Gouty Arthritis in Rats Through Activating NLRP3 Inflammasome and Inflammatory Cytokine Production

    doi: 10.2147/JIR.S351660

    Figure Lengend Snippet: The expression changes of P2X7R on macrophages affect their ability to take up YO-PRO-1. ( A ) At 12h, the expression level of P2X7R in each group of macrophages was detected by flow cytometry. ( B ) Expression of P2X7R among the three groups at different time points. ( C ) At 12h, the percentage of YO-PRO-1 uptake by macrophages in each group was detected by flow cytometry. ( D ) The ability of macrophages to uptake YO-PRO-1 among the three groups at each time point. ( E ) The expression level of P2X7R was positively correlated with the ability of macrophages to uptake YO-PRO-1. * P

    Article Snippet: The treated spleen cells (1×10^6cells) were stained with anti-P2X7 receptor (extracellular)-FITC antibody (Alomone, Jerusalem, Israel) and CD68 antibody (NOVUS, Littleton, USA), and the white cell suspension obtained from the spleen was stained with FITC conjugated mouse anti-rat CD4, PE conjugated mouse anti-rat CD25 (BD Bioscience, MD, USA), PE conjugated anti-IL-17A and APC conjugated anti-Foxp3 antibodies (eBioscience, San Jose, CA, USA).

    Techniques: Expressing, Flow Cytometry

    The changing trend of cytokines and Treg/Th17 cells matched with the pathogenesis process of ATP-activated P2X7R-regulated acute gouty arthritis. ( A ) The trends of P2X7R and IL-1β in ATP group. ( B ) The trends of CD4 + IL-17 + Th17 cells and IL-17 in ATP group. ( C ) The trends of CD4 + CD25 + Foxp3 + Treg cells and TGF-β 1 in ATP group. ( D ) The trends of CD4 + CD25 + Foxp3 + Treg cells and CD4 + IL-17 + Th17 cells in ATP group. ( E ) The ratios of Treg/Th17 among ATP group, BBG group and Control group. * P

    Journal: Journal of Inflammation Research

    Article Title: ATP-Activated P2X7R Promote the Attack of Acute Gouty Arthritis in Rats Through Activating NLRP3 Inflammasome and Inflammatory Cytokine Production

    doi: 10.2147/JIR.S351660

    Figure Lengend Snippet: The changing trend of cytokines and Treg/Th17 cells matched with the pathogenesis process of ATP-activated P2X7R-regulated acute gouty arthritis. ( A ) The trends of P2X7R and IL-1β in ATP group. ( B ) The trends of CD4 + IL-17 + Th17 cells and IL-17 in ATP group. ( C ) The trends of CD4 + CD25 + Foxp3 + Treg cells and TGF-β 1 in ATP group. ( D ) The trends of CD4 + CD25 + Foxp3 + Treg cells and CD4 + IL-17 + Th17 cells in ATP group. ( E ) The ratios of Treg/Th17 among ATP group, BBG group and Control group. * P

    Article Snippet: The treated spleen cells (1×10^6cells) were stained with anti-P2X7 receptor (extracellular)-FITC antibody (Alomone, Jerusalem, Israel) and CD68 antibody (NOVUS, Littleton, USA), and the white cell suspension obtained from the spleen was stained with FITC conjugated mouse anti-rat CD4, PE conjugated mouse anti-rat CD25 (BD Bioscience, MD, USA), PE conjugated anti-IL-17A and APC conjugated anti-Foxp3 antibodies (eBioscience, San Jose, CA, USA).

    Techniques: