p2x7  (Alomone Labs)


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    Structured Review

    Alomone Labs p2x7
    <t>P2X7</t> 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.
    P2x7, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) 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

    2) Product Images from "MicroRNA-22 Controls Aberrant Neurogenesis and Changes in Neuronal Morphology After Status Epilepticus"

    Article Title: MicroRNA-22 Controls Aberrant Neurogenesis and Changes in Neuronal Morphology After Status Epilepticus

    Journal: Frontiers in Molecular Neuroscience

    doi: 10.3389/fnmol.2018.00442

    Increased miR-22 levels in the contralateral hippocampus during epilepsy in mice. (A) Schematic showing intraamygdala injection of kainic acid (KA) into the ipsilateral amygdala to induce status epilepticus (SE). (B) Graph showing increased levels of miR-22 in the contralateral dentate gyrus (DG) 2 weeks following SE ( n = 4 per group, Student’s t -test: * t = 3.483, p = 0.0131). (C) In situ hybridization showing miR-22 in the DG of the contralateral hippocampus in non-epileptic control vehicle-injected mice and in mice subjected to SE. Of note, miR-22 levels seem to be mainly increased in the subgranular zone in the DG of the contralateral hippocampus during epilepsy (6 weeks following SE). Scale bar = 50 μm. (D) MiR-22 expression in the hippocampus in a patient suffering from temporal lobe epilepsy (TLE). MiR-22-positive cells were mainly localized to the DG (arrows and magnification). snRNA U6 was used as positive control. Scale bar = 1 mm. (E) Graph showing decreased miR-22 levels in the hippocampus ( n = 4 per group Student’s t -test: ** t = 4.955, p = 0.0026). (F) In situ hybridization showing decreased miR-22 levels in the subgranular zone of the DG 6 weeks following intra-cerebro-ventricular (i.c.v.) injection of Ant22. Scale bar = 50 μm. (G) Decreased P2X7 receptor expression in the contralateral hippocampus of epileptic mice 14 days following SE when compared to control mice ( n = 4 per group, Students t -test: * t = 2.734, p = 0.034). (H) Lower BzATP-provoked currents detected by patch clamp from green fluorescent protein (GFP)-positive cells present in the contralateral DG of the hippocampus of epileptic P2rx7 -GFP expressing mice when compared to GFP-positive cells from the ipsilateral DG 14 days following SE ( n = 19 cells for ipsi- and 17 cells from contralateral hippocampus from nine mice per group, Student’s t -test, * t = 2.673, p = 0.0115). Scale bar = 50 μm. (I) Increased P2X7 receptor expression mainly localized to the subgranular zone of the DG of the contralateral hippocampus (arrows) in mice treated with Ant22 6 weeks following SE when compared to scramble-treated epileptic mice. n = 5, Scale bar = 50 μm.
    Figure Legend Snippet: Increased miR-22 levels in the contralateral hippocampus during epilepsy in mice. (A) Schematic showing intraamygdala injection of kainic acid (KA) into the ipsilateral amygdala to induce status epilepticus (SE). (B) Graph showing increased levels of miR-22 in the contralateral dentate gyrus (DG) 2 weeks following SE ( n = 4 per group, Student’s t -test: * t = 3.483, p = 0.0131). (C) In situ hybridization showing miR-22 in the DG of the contralateral hippocampus in non-epileptic control vehicle-injected mice and in mice subjected to SE. Of note, miR-22 levels seem to be mainly increased in the subgranular zone in the DG of the contralateral hippocampus during epilepsy (6 weeks following SE). Scale bar = 50 μm. (D) MiR-22 expression in the hippocampus in a patient suffering from temporal lobe epilepsy (TLE). MiR-22-positive cells were mainly localized to the DG (arrows and magnification). snRNA U6 was used as positive control. Scale bar = 1 mm. (E) Graph showing decreased miR-22 levels in the hippocampus ( n = 4 per group Student’s t -test: ** t = 4.955, p = 0.0026). (F) In situ hybridization showing decreased miR-22 levels in the subgranular zone of the DG 6 weeks following intra-cerebro-ventricular (i.c.v.) injection of Ant22. Scale bar = 50 μm. (G) Decreased P2X7 receptor expression in the contralateral hippocampus of epileptic mice 14 days following SE when compared to control mice ( n = 4 per group, Students t -test: * t = 2.734, p = 0.034). (H) Lower BzATP-provoked currents detected by patch clamp from green fluorescent protein (GFP)-positive cells present in the contralateral DG of the hippocampus of epileptic P2rx7 -GFP expressing mice when compared to GFP-positive cells from the ipsilateral DG 14 days following SE ( n = 19 cells for ipsi- and 17 cells from contralateral hippocampus from nine mice per group, Student’s t -test, * t = 2.673, p = 0.0115). Scale bar = 50 μm. (I) Increased P2X7 receptor expression mainly localized to the subgranular zone of the DG of the contralateral hippocampus (arrows) in mice treated with Ant22 6 weeks following SE when compared to scramble-treated epileptic mice. n = 5, Scale bar = 50 μm.

    Techniques Used: Mouse Assay, Injection, In Situ Hybridization, Expressing, Positive Control, Patch Clamp

    3) Product Images from "P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway"

    Article Title: P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway

    Journal: Journal of neurochemistry

    doi: 10.1111/jnc.12286

    Motor neurons in primary culture express P2X7
    Figure Legend Snippet: Motor neurons in primary culture express P2X7

    Techniques Used:

    4) Product Images from "P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway"

    Article Title: P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway

    Journal: Journal of neurochemistry

    doi: 10.1111/jnc.12286

    Motor neurons in primary culture express P2X7
    Figure Legend Snippet: Motor neurons in primary culture express P2X7

    Techniques Used:

    5) Product Images from "P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway"

    Article Title: P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway

    Journal: Journal of neurochemistry

    doi: 10.1111/jnc.12286

    Motor neurons in primary culture express P2X7
    Figure Legend Snippet: Motor neurons in primary culture express P2X7

    Techniques Used:

    6) Product Images from "Staphylococcus aureus Activates the NLRP3 Inflammasome in Human and Rat Conjunctival Goblet Cells"

    Article Title: Staphylococcus aureus Activates the NLRP3 Inflammasome in Human and Rat Conjunctival Goblet Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0074010

    Purinergic receptors P2X4, P2X7, and TLR2 are expressed in the rat conjunctiva and rat goblet cell cultures. All three receptors were identified by red immunofluorescent staining. UEA stains goblet cell secretory products green, allowing the identification of goblet cells within the conjunctiva. Rabbit isotype controls were negative. Arrows indicate location of goblet cells.
    Figure Legend Snippet: Purinergic receptors P2X4, P2X7, and TLR2 are expressed in the rat conjunctiva and rat goblet cell cultures. All three receptors were identified by red immunofluorescent staining. UEA stains goblet cell secretory products green, allowing the identification of goblet cells within the conjunctiva. Rabbit isotype controls were negative. Arrows indicate location of goblet cells.

    Techniques Used: Staining

    The purinergic receptors P2X4, P2X7, and TLR2 are functional in cultured rat goblet cells. Rat goblet cells were loaded with fura-2 and then stimulated with ATP and intracellular calcium response measured. A typical trace from one experiment with ATP (5 mM, A ) or lipoteichoic acid (LTA, 10 µg/ml, B ) is representative of 3 animals. Representative photographs of a single field of cells after treatment with ATP or LTA, with warmer colors indicating intracellular Ca 2+ increase, are shown as insets. Peak [Ca 2+ ] i calculated for ATP (0.1 µM–5 mM) from 3 experiments, is shown in C . Peak [Ca 2+ ] i for goblet cells, also preincubated with LTA for 5 h and then loaded with fura-2 for 1 additional h before addition of ATP (5 mM), calculated from 3 experiments, is shown in D . Results are expressed as mean ± SEM. * indicates significance of p
    Figure Legend Snippet: The purinergic receptors P2X4, P2X7, and TLR2 are functional in cultured rat goblet cells. Rat goblet cells were loaded with fura-2 and then stimulated with ATP and intracellular calcium response measured. A typical trace from one experiment with ATP (5 mM, A ) or lipoteichoic acid (LTA, 10 µg/ml, B ) is representative of 3 animals. Representative photographs of a single field of cells after treatment with ATP or LTA, with warmer colors indicating intracellular Ca 2+ increase, are shown as insets. Peak [Ca 2+ ] i calculated for ATP (0.1 µM–5 mM) from 3 experiments, is shown in C . Peak [Ca 2+ ] i for goblet cells, also preincubated with LTA for 5 h and then loaded with fura-2 for 1 additional h before addition of ATP (5 mM), calculated from 3 experiments, is shown in D . Results are expressed as mean ± SEM. * indicates significance of p

    Techniques Used: Functional Assay, Cell Culture

    Purinergic receptors P2X4, P2X7 and TLR2 are expressed in rat goblet cell cultures. All three receptors were identified by western blot. Lanes 1–3 represent separate animals. B is the positive control of rat brain. The β-actin blot is for both P2X4 and P2X7 receptors blots.
    Figure Legend Snippet: Purinergic receptors P2X4, P2X7 and TLR2 are expressed in rat goblet cell cultures. All three receptors were identified by western blot. Lanes 1–3 represent separate animals. B is the positive control of rat brain. The β-actin blot is for both P2X4 and P2X7 receptors blots.

    Techniques Used: Western Blot, Positive Control

    7) 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

    8) 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

    9) Product Images from "MicroRNA-22 Controls Aberrant Neurogenesis and Changes in Neuronal Morphology After Status Epilepticus"

    Article Title: MicroRNA-22 Controls Aberrant Neurogenesis and Changes in Neuronal Morphology After Status Epilepticus

    Journal: Frontiers in Molecular Neuroscience

    doi: 10.3389/fnmol.2018.00442

    Increased miR-22 levels in the contralateral hippocampus during epilepsy in mice. (A) Schematic showing intraamygdala injection of kainic acid (KA) into the ipsilateral amygdala to induce status epilepticus (SE). (B) Graph showing increased levels of miR-22 in the contralateral dentate gyrus (DG) 2 weeks following SE ( n = 4 per group, Student’s t -test: * t = 3.483, p = 0.0131). (C) In situ hybridization showing miR-22 in the DG of the contralateral hippocampus in non-epileptic control vehicle-injected mice and in mice subjected to SE. Of note, miR-22 levels seem to be mainly increased in the subgranular zone in the DG of the contralateral hippocampus during epilepsy (6 weeks following SE). Scale bar = 50 μm. (D) MiR-22 expression in the hippocampus in a patient suffering from temporal lobe epilepsy (TLE). MiR-22-positive cells were mainly localized to the DG (arrows and magnification). snRNA U6 was used as positive control. Scale bar = 1 mm. (E) Graph showing decreased miR-22 levels in the hippocampus ( n = 4 per group Student’s t -test: ** t = 4.955, p = 0.0026). (F) In situ hybridization showing decreased miR-22 levels in the subgranular zone of the DG 6 weeks following intra-cerebro-ventricular (i.c.v.) injection of Ant22. Scale bar = 50 μm. (G) Decreased P2X7 receptor expression in the contralateral hippocampus of epileptic mice 14 days following SE when compared to control mice ( n = 4 per group, Students t -test: * t = 2.734, p = 0.034). (H) Lower BzATP-provoked currents detected by patch clamp from green fluorescent protein (GFP)-positive cells present in the contralateral DG of the hippocampus of epileptic P2rx7 -GFP expressing mice when compared to GFP-positive cells from the ipsilateral DG 14 days following SE ( n = 19 cells for ipsi- and 17 cells from contralateral hippocampus from nine mice per group, Student’s t -test, * t = 2.673, p = 0.0115). Scale bar = 50 μm. (I) Increased P2X7 receptor expression mainly localized to the subgranular zone of the DG of the contralateral hippocampus (arrows) in mice treated with Ant22 6 weeks following SE when compared to scramble-treated epileptic mice. n = 5, Scale bar = 50 μm.
    Figure Legend Snippet: Increased miR-22 levels in the contralateral hippocampus during epilepsy in mice. (A) Schematic showing intraamygdala injection of kainic acid (KA) into the ipsilateral amygdala to induce status epilepticus (SE). (B) Graph showing increased levels of miR-22 in the contralateral dentate gyrus (DG) 2 weeks following SE ( n = 4 per group, Student’s t -test: * t = 3.483, p = 0.0131). (C) In situ hybridization showing miR-22 in the DG of the contralateral hippocampus in non-epileptic control vehicle-injected mice and in mice subjected to SE. Of note, miR-22 levels seem to be mainly increased in the subgranular zone in the DG of the contralateral hippocampus during epilepsy (6 weeks following SE). Scale bar = 50 μm. (D) MiR-22 expression in the hippocampus in a patient suffering from temporal lobe epilepsy (TLE). MiR-22-positive cells were mainly localized to the DG (arrows and magnification). snRNA U6 was used as positive control. Scale bar = 1 mm. (E) Graph showing decreased miR-22 levels in the hippocampus ( n = 4 per group Student’s t -test: ** t = 4.955, p = 0.0026). (F) In situ hybridization showing decreased miR-22 levels in the subgranular zone of the DG 6 weeks following intra-cerebro-ventricular (i.c.v.) injection of Ant22. Scale bar = 50 μm. (G) Decreased P2X7 receptor expression in the contralateral hippocampus of epileptic mice 14 days following SE when compared to control mice ( n = 4 per group, Students t -test: * t = 2.734, p = 0.034). (H) Lower BzATP-provoked currents detected by patch clamp from green fluorescent protein (GFP)-positive cells present in the contralateral DG of the hippocampus of epileptic P2rx7 -GFP expressing mice when compared to GFP-positive cells from the ipsilateral DG 14 days following SE ( n = 19 cells for ipsi- and 17 cells from contralateral hippocampus from nine mice per group, Student’s t -test, * t = 2.673, p = 0.0115). Scale bar = 50 μm. (I) Increased P2X7 receptor expression mainly localized to the subgranular zone of the DG of the contralateral hippocampus (arrows) in mice treated with Ant22 6 weeks following SE when compared to scramble-treated epileptic mice. n = 5, Scale bar = 50 μm.

    Techniques Used: Mouse Assay, Injection, In Situ Hybridization, Expressing, Positive Control, Patch Clamp

    10) Product Images from "Synthetic cathinone MDPV enhances reward function through purinergic P2X7 receptor-dependent pathway and increases P2X7 gene expression in nucleus accumbens"

    Article Title: Synthetic cathinone MDPV enhances reward function through purinergic P2X7 receptor-dependent pathway and increases P2X7 gene expression in nucleus accumbens

    Journal: Drug and alcohol dependence

    doi: 10.1016/j.drugalcdep.2018.12.022

    P2X7 antagonist A438079 inhibits reduction in brain reward threshold produced by MDPV. Percentage change in baseline reward threshold + S.E.M. shown in (A, C, E) and maximum response rate shown in (B, D, F). (A-B) MDPV (0.5 mg/kg) reduces brain reward threshold without affecting maximal response rate. N=4-7. ** p
    Figure Legend Snippet: P2X7 antagonist A438079 inhibits reduction in brain reward threshold produced by MDPV. Percentage change in baseline reward threshold + S.E.M. shown in (A, C, E) and maximum response rate shown in (B, D, F). (A-B) MDPV (0.5 mg/kg) reduces brain reward threshold without affecting maximal response rate. N=4-7. ** p

    Techniques Used: Produced

    11) Product Images from "P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway"

    Article Title: P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway

    Journal: Journal of neurochemistry

    doi: 10.1111/jnc.12286

    Motor neurons in primary culture express P2X7
    Figure Legend Snippet: Motor neurons in primary culture express P2X7

    Techniques Used:

    12) 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

    13) Product Images from "MicroRNA-22 Controls Aberrant Neurogenesis and Changes in Neuronal Morphology After Status Epilepticus"

    Article Title: MicroRNA-22 Controls Aberrant Neurogenesis and Changes in Neuronal Morphology After Status Epilepticus

    Journal: Frontiers in Molecular Neuroscience

    doi: 10.3389/fnmol.2018.00442

    Increased miR-22 levels in the contralateral hippocampus during epilepsy in mice. (A) Schematic showing intraamygdala injection of kainic acid (KA) into the ipsilateral amygdala to induce status epilepticus (SE). (B) Graph showing increased levels of miR-22 in the contralateral dentate gyrus (DG) 2 weeks following SE ( n = 4 per group, Student’s t -test: * t = 3.483, p = 0.0131). (C) In situ hybridization showing miR-22 in the DG of the contralateral hippocampus in non-epileptic control vehicle-injected mice and in mice subjected to SE. Of note, miR-22 levels seem to be mainly increased in the subgranular zone in the DG of the contralateral hippocampus during epilepsy (6 weeks following SE). Scale bar = 50 μm. (D) MiR-22 expression in the hippocampus in a patient suffering from temporal lobe epilepsy (TLE). MiR-22-positive cells were mainly localized to the DG (arrows and magnification). snRNA U6 was used as positive control. Scale bar = 1 mm. (E) Graph showing decreased miR-22 levels in the hippocampus ( n = 4 per group Student’s t -test: ** t = 4.955, p = 0.0026). (F) In situ hybridization showing decreased miR-22 levels in the subgranular zone of the DG 6 weeks following intra-cerebro-ventricular (i.c.v.) injection of Ant22. Scale bar = 50 μm. (G) Decreased P2X7 receptor expression in the contralateral hippocampus of epileptic mice 14 days following SE when compared to control mice ( n = 4 per group, Students t -test: * t = 2.734, p = 0.034). (H) Lower BzATP-provoked currents detected by patch clamp from green fluorescent protein (GFP)-positive cells present in the contralateral DG of the hippocampus of epileptic P2rx7 -GFP expressing mice when compared to GFP-positive cells from the ipsilateral DG 14 days following SE ( n = 19 cells for ipsi- and 17 cells from contralateral hippocampus from nine mice per group, Student’s t -test, * t = 2.673, p = 0.0115). Scale bar = 50 μm. (I) Increased P2X7 receptor expression mainly localized to the subgranular zone of the DG of the contralateral hippocampus (arrows) in mice treated with Ant22 6 weeks following SE when compared to scramble-treated epileptic mice. n = 5, Scale bar = 50 μm.
    Figure Legend Snippet: Increased miR-22 levels in the contralateral hippocampus during epilepsy in mice. (A) Schematic showing intraamygdala injection of kainic acid (KA) into the ipsilateral amygdala to induce status epilepticus (SE). (B) Graph showing increased levels of miR-22 in the contralateral dentate gyrus (DG) 2 weeks following SE ( n = 4 per group, Student’s t -test: * t = 3.483, p = 0.0131). (C) In situ hybridization showing miR-22 in the DG of the contralateral hippocampus in non-epileptic control vehicle-injected mice and in mice subjected to SE. Of note, miR-22 levels seem to be mainly increased in the subgranular zone in the DG of the contralateral hippocampus during epilepsy (6 weeks following SE). Scale bar = 50 μm. (D) MiR-22 expression in the hippocampus in a patient suffering from temporal lobe epilepsy (TLE). MiR-22-positive cells were mainly localized to the DG (arrows and magnification). snRNA U6 was used as positive control. Scale bar = 1 mm. (E) Graph showing decreased miR-22 levels in the hippocampus ( n = 4 per group Student’s t -test: ** t = 4.955, p = 0.0026). (F) In situ hybridization showing decreased miR-22 levels in the subgranular zone of the DG 6 weeks following intra-cerebro-ventricular (i.c.v.) injection of Ant22. Scale bar = 50 μm. (G) Decreased P2X7 receptor expression in the contralateral hippocampus of epileptic mice 14 days following SE when compared to control mice ( n = 4 per group, Students t -test: * t = 2.734, p = 0.034). (H) Lower BzATP-provoked currents detected by patch clamp from green fluorescent protein (GFP)-positive cells present in the contralateral DG of the hippocampus of epileptic P2rx7 -GFP expressing mice when compared to GFP-positive cells from the ipsilateral DG 14 days following SE ( n = 19 cells for ipsi- and 17 cells from contralateral hippocampus from nine mice per group, Student’s t -test, * t = 2.673, p = 0.0115). Scale bar = 50 μm. (I) Increased P2X7 receptor expression mainly localized to the subgranular zone of the DG of the contralateral hippocampus (arrows) in mice treated with Ant22 6 weeks following SE when compared to scramble-treated epileptic mice. n = 5, Scale bar = 50 μm.

    Techniques Used: Mouse Assay, Injection, In Situ Hybridization, Expressing, Positive Control, Patch Clamp

    14) Product Images from "Staphylococcus aureus Activates the NLRP3 Inflammasome in Human and Rat Conjunctival Goblet Cells"

    Article Title: Staphylococcus aureus Activates the NLRP3 Inflammasome in Human and Rat Conjunctival Goblet Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0074010

    Purinergic receptors P2X4, P2X7, and TLR2 are expressed in the rat conjunctiva and rat goblet cell cultures. All three receptors were identified by red immunofluorescent staining. UEA stains goblet cell secretory products green, allowing the identification of goblet cells within the conjunctiva. Rabbit isotype controls were negative. Arrows indicate location of goblet cells.
    Figure Legend Snippet: Purinergic receptors P2X4, P2X7, and TLR2 are expressed in the rat conjunctiva and rat goblet cell cultures. All three receptors were identified by red immunofluorescent staining. UEA stains goblet cell secretory products green, allowing the identification of goblet cells within the conjunctiva. Rabbit isotype controls were negative. Arrows indicate location of goblet cells.

    Techniques Used: Staining

    The purinergic receptors P2X4, P2X7, and TLR2 are functional in cultured rat goblet cells. Rat goblet cells were loaded with fura-2 and then stimulated with ATP and intracellular calcium response measured. A typical trace from one experiment with ATP (5 mM, A ) or lipoteichoic acid (LTA, 10 µg/ml, B ) is representative of 3 animals. Representative photographs of a single field of cells after treatment with ATP or LTA, with warmer colors indicating intracellular Ca 2+ increase, are shown as insets. Peak [Ca 2+ ] i calculated for ATP (0.1 µM–5 mM) from 3 experiments, is shown in C . Peak [Ca 2+ ] i for goblet cells, also preincubated with LTA for 5 h and then loaded with fura-2 for 1 additional h before addition of ATP (5 mM), calculated from 3 experiments, is shown in D . Results are expressed as mean ± SEM. * indicates significance of p
    Figure Legend Snippet: The purinergic receptors P2X4, P2X7, and TLR2 are functional in cultured rat goblet cells. Rat goblet cells were loaded with fura-2 and then stimulated with ATP and intracellular calcium response measured. A typical trace from one experiment with ATP (5 mM, A ) or lipoteichoic acid (LTA, 10 µg/ml, B ) is representative of 3 animals. Representative photographs of a single field of cells after treatment with ATP or LTA, with warmer colors indicating intracellular Ca 2+ increase, are shown as insets. Peak [Ca 2+ ] i calculated for ATP (0.1 µM–5 mM) from 3 experiments, is shown in C . Peak [Ca 2+ ] i for goblet cells, also preincubated with LTA for 5 h and then loaded with fura-2 for 1 additional h before addition of ATP (5 mM), calculated from 3 experiments, is shown in D . Results are expressed as mean ± SEM. * indicates significance of p

    Techniques Used: Functional Assay, Cell Culture

    Purinergic receptors P2X4, P2X7 and TLR2 are expressed in rat goblet cell cultures. All three receptors were identified by western blot. Lanes 1–3 represent separate animals. B is the positive control of rat brain. The β-actin blot is for both P2X4 and P2X7 receptors blots.
    Figure Legend Snippet: Purinergic receptors P2X4, P2X7 and TLR2 are expressed in rat goblet cell cultures. All three receptors were identified by western blot. Lanes 1–3 represent separate animals. B is the positive control of rat brain. The β-actin blot is for both P2X4 and P2X7 receptors blots.

    Techniques Used: Western Blot, Positive Control

    15) Product Images from "P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway"

    Article Title: P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway

    Journal: Journal of neurochemistry

    doi: 10.1111/jnc.12286

    Motor neurons in primary culture express P2X7
    Figure Legend Snippet: Motor neurons in primary culture express P2X7

    Techniques Used:

    16) Product Images from "P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway"

    Article Title: P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway

    Journal: Journal of neurochemistry

    doi: 10.1111/jnc.12286

    Motor neurons in primary culture express P2X7
    Figure Legend Snippet: Motor neurons in primary culture express P2X7

    Techniques Used:

    17) Product Images from "P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway"

    Article Title: P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway

    Journal: Journal of neurochemistry

    doi: 10.1111/jnc.12286

    Motor neurons in primary culture express P2X7
    Figure Legend Snippet: Motor neurons in primary culture express P2X7

    Techniques Used:

    18) Product Images from "P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway"

    Article Title: P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway

    Journal: Journal of neurochemistry

    doi: 10.1111/jnc.12286

    Motor neurons in primary culture express P2X7
    Figure Legend Snippet: Motor neurons in primary culture express P2X7

    Techniques Used:

    19) 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

    20) Product Images from "P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway"

    Article Title: P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway

    Journal: Journal of neurochemistry

    doi: 10.1111/jnc.12286

    Motor neurons in primary culture express P2X7
    Figure Legend Snippet: Motor neurons in primary culture express P2X7

    Techniques Used:

    21) Product Images from "P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway"

    Article Title: P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway

    Journal: Journal of neurochemistry

    doi: 10.1111/jnc.12286

    Motor neurons in primary culture express P2X7
    Figure Legend Snippet: Motor neurons in primary culture express P2X7

    Techniques Used:

    22) 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

    23) Product Images from "P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway"

    Article Title: P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway

    Journal: Journal of neurochemistry

    doi: 10.1111/jnc.12286

    Motor neurons in primary culture express P2X7
    Figure Legend Snippet: Motor neurons in primary culture express P2X7

    Techniques Used:

    24) Product Images from "Staphylococcus aureus Activates the NLRP3 Inflammasome in Human and Rat Conjunctival Goblet Cells"

    Article Title: Staphylococcus aureus Activates the NLRP3 Inflammasome in Human and Rat Conjunctival Goblet Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0074010

    Purinergic receptors P2X4, P2X7, and TLR2 are expressed in the rat conjunctiva and rat goblet cell cultures. All three receptors were identified by red immunofluorescent staining. UEA stains goblet cell secretory products green, allowing the identification of goblet cells within the conjunctiva. Rabbit isotype controls were negative. Arrows indicate location of goblet cells.
    Figure Legend Snippet: Purinergic receptors P2X4, P2X7, and TLR2 are expressed in the rat conjunctiva and rat goblet cell cultures. All three receptors were identified by red immunofluorescent staining. UEA stains goblet cell secretory products green, allowing the identification of goblet cells within the conjunctiva. Rabbit isotype controls were negative. Arrows indicate location of goblet cells.

    Techniques Used: Staining

    The purinergic receptors P2X4, P2X7, and TLR2 are functional in cultured rat goblet cells. Rat goblet cells were loaded with fura-2 and then stimulated with ATP and intracellular calcium response measured. A typical trace from one experiment with ATP (5 mM, A ) or lipoteichoic acid (LTA, 10 µg/ml, B ) is representative of 3 animals. Representative photographs of a single field of cells after treatment with ATP or LTA, with warmer colors indicating intracellular Ca 2+ increase, are shown as insets. Peak [Ca 2+ ] i calculated for ATP (0.1 µM–5 mM) from 3 experiments, is shown in C . Peak [Ca 2+ ] i for goblet cells, also preincubated with LTA for 5 h and then loaded with fura-2 for 1 additional h before addition of ATP (5 mM), calculated from 3 experiments, is shown in D . Results are expressed as mean ± SEM. * indicates significance of p
    Figure Legend Snippet: The purinergic receptors P2X4, P2X7, and TLR2 are functional in cultured rat goblet cells. Rat goblet cells were loaded with fura-2 and then stimulated with ATP and intracellular calcium response measured. A typical trace from one experiment with ATP (5 mM, A ) or lipoteichoic acid (LTA, 10 µg/ml, B ) is representative of 3 animals. Representative photographs of a single field of cells after treatment with ATP or LTA, with warmer colors indicating intracellular Ca 2+ increase, are shown as insets. Peak [Ca 2+ ] i calculated for ATP (0.1 µM–5 mM) from 3 experiments, is shown in C . Peak [Ca 2+ ] i for goblet cells, also preincubated with LTA for 5 h and then loaded with fura-2 for 1 additional h before addition of ATP (5 mM), calculated from 3 experiments, is shown in D . Results are expressed as mean ± SEM. * indicates significance of p

    Techniques Used: Functional Assay, Cell Culture

    Purinergic receptors P2X4, P2X7 and TLR2 are expressed in rat goblet cell cultures. All three receptors were identified by western blot. Lanes 1–3 represent separate animals. B is the positive control of rat brain. The β-actin blot is for both P2X4 and P2X7 receptors blots.
    Figure Legend Snippet: Purinergic receptors P2X4, P2X7 and TLR2 are expressed in rat goblet cell cultures. All three receptors were identified by western blot. Lanes 1–3 represent separate animals. B is the positive control of rat brain. The β-actin blot is for both P2X4 and P2X7 receptors blots.

    Techniques Used: Western Blot, Positive Control

    25) 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

    26) Product Images from "P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway"

    Article Title: P2X7 receptor-induced death of motor neurons by a peroxynitrite/FAS-dependent pathway

    Journal: Journal of neurochemistry

    doi: 10.1111/jnc.12286

    Motor neurons in primary culture express P2X7
    Figure Legend Snippet: Motor neurons in primary culture express P2X7

    Techniques Used:

    27) 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

    28) 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

    29) 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

    30) 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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  • 88
    Alomone Labs rabbit polyclonal anti human p2x7 receptor atto 488
    TLR4 signaling influences <t>P2X7</t> receptor distribution. Representative confocal microphotographs showing (A) HuC/D (magenta) and P2X7Rs (yellow) distribution and (B) analysis of P2X7Rs fluorescence intensities in WT and TLR4 -/- preparations ( N = 5 mice/group). Cell nuclei were stained with TOTO-3 (blue). Bars = 22 μm. ∗ P
    Rabbit Polyclonal Anti Human P2x7 Receptor Atto 488, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti human p2x7 receptor atto 488/product/Alomone Labs
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    rabbit polyclonal anti human p2x7 receptor atto 488 - by Bioz Stars, 2022-10
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    86
    Alomone Labs rabbit polyclonal p2x7 c terminus antibody
    The N terminus is important for the sensitivity of P2X7 receptor activity to cholesterol depletion. A , amino acid sequence alignment of the N-terminal region of human P2X1, P2X2, and P2X7 receptors, with conserved amino acids highlighted in  gray , P2X1 aa20–23 and equivalent residues in P2X2  underlined , and the P2X7 residues Lys 17  and Val 18  in  red. B , time course of dye uptake mediated by the K17R/V18M mutant and wild type P2X7, alone, pretreated with MCD and loaded with cholesterol ( chol ), normalized to the untreated WT P2X7.  C , bar graph of the relative rate of dye uptake normalized to the untreated WT P2X7. Dye uptake mediated by the K17R/V18M mutant is not potentiated by MCD, but is inhibited by cholesterol loading (WT:  n  = 14 alone,  n  = 11 MCD,  n  = 3 cholesterol; mutants:  n  = 12 alone,  n  = 11 MCD,  n  = 3 cholesterol; *,  p
    Rabbit Polyclonal P2x7 C Terminus Antibody, 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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    Alomone Labs p2x7r
    Activation of the <t>P2X7R</t> induces APP cleavage by a metalloprotease. Neuro2a-hAPP cells preincubated for 1 h at 37 °C with 50 μ m Z-VAD-FMK ( A and B ) or increasing concentrations of GM6001 ( C and D ) or TAPI-2 ( E and F ), and stimulated with
    P2x7r, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs p2x7
    <t>P2X7</t> 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.
    P2x7, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TLR4 signaling influences P2X7 receptor distribution. Representative confocal microphotographs showing (A) HuC/D (magenta) and P2X7Rs (yellow) distribution and (B) analysis of P2X7Rs fluorescence intensities in WT and TLR4 -/- preparations ( N = 5 mice/group). Cell nuclei were stained with TOTO-3 (blue). Bars = 22 μm. ∗ P

    Journal: Frontiers in Pharmacology

    Article Title: Toll-Like Receptor 4 Modulates Small Intestine Neuromuscular Function through Nitrergic and Purinergic Pathways

    doi: 10.3389/fphar.2017.00350

    Figure Lengend Snippet: TLR4 signaling influences P2X7 receptor distribution. Representative confocal microphotographs showing (A) HuC/D (magenta) and P2X7Rs (yellow) distribution and (B) analysis of P2X7Rs fluorescence intensities in WT and TLR4 -/- preparations ( N = 5 mice/group). Cell nuclei were stained with TOTO-3 (blue). Bars = 22 μm. ∗ P

    Article Snippet: LMMP whole mount preparations or ileal cryosections were then incubated overnight at room temperature with the following antibodies: chicken polyclonal anti-mouse glial fibrillary acidic protein (GFAP; 1:100; Abcam, Milan, Italy), rabbit polyclonal anti-human GFAP (1:200; Merck Millipore Corporation, Milan, Italy), mouse biotin-conjugated anti-human HuC/D (1:50; Thermo Fisher Scientific, Milan, Italy), rabbit polyclonal anti-mouse neuronal nitric oxide synthase (nNOS, 1:100; Thermo Fisher Scientific, Milan, Italy), rabbit polyclonal anti-human inducible NOS (iNOS; 1:50, Santa Cruz Biotechnology, Milan, Italy), rabbit polyclonal anti-human vasoactive intestinal peptide (VIP, 1:100; GenWay Biotech, Milan, Italy), rabbit polyclonal anti-human P2X7 receptor-ATTO-488 (1:100; Alomone labs, Jerusalem, Israel), rabbit polyclonal anti-human P2Y1 receptor (1:100; Alomone labs, Jerusalem, Israel), rabbit monoclonal anti-human S100β (1:100; Merck Millipore Corporation, Milan, Italy) and guinea pig polyclonal anti-mouse substance P (1:100; Abcam, Milan, Italy).

    Techniques: Fluorescence, Mouse Assay, Staining

    The N terminus is important for the sensitivity of P2X7 receptor activity to cholesterol depletion. A , amino acid sequence alignment of the N-terminal region of human P2X1, P2X2, and P2X7 receptors, with conserved amino acids highlighted in  gray , P2X1 aa20–23 and equivalent residues in P2X2  underlined , and the P2X7 residues Lys 17  and Val 18  in  red. B , time course of dye uptake mediated by the K17R/V18M mutant and wild type P2X7, alone, pretreated with MCD and loaded with cholesterol ( chol ), normalized to the untreated WT P2X7.  C , bar graph of the relative rate of dye uptake normalized to the untreated WT P2X7. Dye uptake mediated by the K17R/V18M mutant is not potentiated by MCD, but is inhibited by cholesterol loading (WT:  n  = 14 alone,  n  = 11 MCD,  n  = 3 cholesterol; mutants:  n  = 12 alone,  n  = 11 MCD,  n  = 3 cholesterol; *,  p

    Journal: The Journal of Biological Chemistry

    Article Title: Plasma Membrane Cholesterol as a Regulator of Human and Rodent P2X7 Receptor Activation and Sensitization *

    doi: 10.1074/jbc.M114.574699

    Figure Lengend Snippet: The N terminus is important for the sensitivity of P2X7 receptor activity to cholesterol depletion. A , amino acid sequence alignment of the N-terminal region of human P2X1, P2X2, and P2X7 receptors, with conserved amino acids highlighted in gray , P2X1 aa20–23 and equivalent residues in P2X2 underlined , and the P2X7 residues Lys 17 and Val 18 in red. B , time course of dye uptake mediated by the K17R/V18M mutant and wild type P2X7, alone, pretreated with MCD and loaded with cholesterol ( chol ), normalized to the untreated WT P2X7. C , bar graph of the relative rate of dye uptake normalized to the untreated WT P2X7. Dye uptake mediated by the K17R/V18M mutant is not potentiated by MCD, but is inhibited by cholesterol loading (WT: n = 14 alone, n = 11 MCD, n = 3 cholesterol; mutants: n = 12 alone, n = 11 MCD, n = 3 cholesterol; *, p

    Article Snippet: Rabbit polyclonal P2X7 C terminus antibody was obtained from Alomone Labs (Jerusalem, Israel), and anti-mouse and anti-rabbit HRP-conjugated secondary antibodies were from Thermo Fisher Scientific and Bio-Rad, respectively.

    Techniques: Activity Assay, Sequencing, Mutagenesis, Relative Rate

    Activation of the P2X7R induces APP cleavage by a metalloprotease. Neuro2a-hAPP cells preincubated for 1 h at 37 °C with 50 μ m Z-VAD-FMK ( A and B ) or increasing concentrations of GM6001 ( C and D ) or TAPI-2 ( E and F ), and stimulated with

    Journal: The Journal of Biological Chemistry

    Article Title: The Purinergic Receptor P2X7 Triggers ?-Secretase-dependent Processing of the Amyloid Precursor Protein *

    doi: 10.1074/jbc.M110.200618

    Figure Lengend Snippet: Activation of the P2X7R induces APP cleavage by a metalloprotease. Neuro2a-hAPP cells preincubated for 1 h at 37 °C with 50 μ m Z-VAD-FMK ( A and B ) or increasing concentrations of GM6001 ( C and D ) or TAPI-2 ( E and F ), and stimulated with

    Article Snippet: Several lines of evidence demonstrate that ATP- or Bz-ATP-mediated sAPPα release is specifically dependent on P2X7R: ( ) three pharmacological inhibitors of P2X7R block the release of sAPPα mediated by Bz-ATP; ( ) inhibition of P2X7R synthesis by RNA interference results in 68.5 ± 7.1% reduction of sAPPα shedding ( , D and E ); and ( ) stimulation by Bz-ATP of mouse primary astrocytes and NPCs from P2X7R-deficient mice does not induce sAPPα release while it does in cells derived from C57BL/6 animals ( , F and G ).

    Techniques: Activation Assay

    P2X7R-dependent extracellular Ca 2+ influx is not involved in sAPP release. A , Neuro2a-hAPP cells were loaded with 2 μ m Fura-2 AM for 30 min. Then, these cells were stimulated with 300 μ m Bz-ATP (■) or 1 μ m thapsigargin

    Journal: The Journal of Biological Chemistry

    Article Title: The Purinergic Receptor P2X7 Triggers ?-Secretase-dependent Processing of the Amyloid Precursor Protein *

    doi: 10.1074/jbc.M110.200618

    Figure Lengend Snippet: P2X7R-dependent extracellular Ca 2+ influx is not involved in sAPP release. A , Neuro2a-hAPP cells were loaded with 2 μ m Fura-2 AM for 30 min. Then, these cells were stimulated with 300 μ m Bz-ATP (■) or 1 μ m thapsigargin

    Article Snippet: Several lines of evidence demonstrate that ATP- or Bz-ATP-mediated sAPPα release is specifically dependent on P2X7R: ( ) three pharmacological inhibitors of P2X7R block the release of sAPPα mediated by Bz-ATP; ( ) inhibition of P2X7R synthesis by RNA interference results in 68.5 ± 7.1% reduction of sAPPα shedding ( , D and E ); and ( ) stimulation by Bz-ATP of mouse primary astrocytes and NPCs from P2X7R-deficient mice does not induce sAPPα release while it does in cells derived from C57BL/6 animals ( , F and G ).

    Techniques:

    Role of MAP kinase pathways in P2X7R-dependent α-cleavage of APP. A , a kinetic study of Erk1/2 phosphorylation after 1 m m Bz-ATP stimulation is shown in the upper left Western blot. The effect of MEK inhibitor is shown on the upper right and

    Journal: The Journal of Biological Chemistry

    Article Title: The Purinergic Receptor P2X7 Triggers ?-Secretase-dependent Processing of the Amyloid Precursor Protein *

    doi: 10.1074/jbc.M110.200618

    Figure Lengend Snippet: Role of MAP kinase pathways in P2X7R-dependent α-cleavage of APP. A , a kinetic study of Erk1/2 phosphorylation after 1 m m Bz-ATP stimulation is shown in the upper left Western blot. The effect of MEK inhibitor is shown on the upper right and

    Article Snippet: Several lines of evidence demonstrate that ATP- or Bz-ATP-mediated sAPPα release is specifically dependent on P2X7R: ( ) three pharmacological inhibitors of P2X7R block the release of sAPPα mediated by Bz-ATP; ( ) inhibition of P2X7R synthesis by RNA interference results in 68.5 ± 7.1% reduction of sAPPα shedding ( , D and E ); and ( ) stimulation by Bz-ATP of mouse primary astrocytes and NPCs from P2X7R-deficient mice does not induce sAPPα release while it does in cells derived from C57BL/6 animals ( , F and G ).

    Techniques: Western Blot

    P2X7R-mediated APP processing is independent of ADAM9, ADAM10, and ADAM17. A , cell lysates of Neuro2a-hAPP cells transfected with control, ADAM10 or ADAM17 siRNA were analyzed by Western blot using anti-ADAM10 or anti-ADAM17 Abs. B , Neuro2a-hAPP cells

    Journal: The Journal of Biological Chemistry

    Article Title: The Purinergic Receptor P2X7 Triggers ?-Secretase-dependent Processing of the Amyloid Precursor Protein *

    doi: 10.1074/jbc.M110.200618

    Figure Lengend Snippet: P2X7R-mediated APP processing is independent of ADAM9, ADAM10, and ADAM17. A , cell lysates of Neuro2a-hAPP cells transfected with control, ADAM10 or ADAM17 siRNA were analyzed by Western blot using anti-ADAM10 or anti-ADAM17 Abs. B , Neuro2a-hAPP cells

    Article Snippet: Several lines of evidence demonstrate that ATP- or Bz-ATP-mediated sAPPα release is specifically dependent on P2X7R: ( ) three pharmacological inhibitors of P2X7R block the release of sAPPα mediated by Bz-ATP; ( ) inhibition of P2X7R synthesis by RNA interference results in 68.5 ± 7.1% reduction of sAPPα shedding ( , D and E ); and ( ) stimulation by Bz-ATP of mouse primary astrocytes and NPCs from P2X7R-deficient mice does not induce sAPPα release while it does in cells derived from C57BL/6 animals ( , F and G ).

    Techniques: Transfection, Western Blot

    Bz-ATP stimulation induces sAPP release via activation of P2X7R. Neuro2a-hAPP cells were preincubated at 37 °C for 1 h with 10 μ m and 50 μ m BBG ( A ), for 2 h with 300 μ m oATP ( B ), and for 30 min with 10 μ m A438079

    Journal: The Journal of Biological Chemistry

    Article Title: The Purinergic Receptor P2X7 Triggers ?-Secretase-dependent Processing of the Amyloid Precursor Protein *

    doi: 10.1074/jbc.M110.200618

    Figure Lengend Snippet: Bz-ATP stimulation induces sAPP release via activation of P2X7R. Neuro2a-hAPP cells were preincubated at 37 °C for 1 h with 10 μ m and 50 μ m BBG ( A ), for 2 h with 300 μ m oATP ( B ), and for 30 min with 10 μ m A438079

    Article Snippet: Several lines of evidence demonstrate that ATP- or Bz-ATP-mediated sAPPα release is specifically dependent on P2X7R: ( ) three pharmacological inhibitors of P2X7R block the release of sAPPα mediated by Bz-ATP; ( ) inhibition of P2X7R synthesis by RNA interference results in 68.5 ± 7.1% reduction of sAPPα shedding ( , D and E ); and ( ) stimulation by Bz-ATP of mouse primary astrocytes and NPCs from P2X7R-deficient mice does not induce sAPPα release while it does in cells derived from C57BL/6 animals ( , F and G ).

    Techniques: Activation Assay

    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.

    Journal: Cardiovascular Research

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

    doi: 10.1093/cvr/cvx213

    Figure Lengend 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.

    Article Snippet: These effects were specific to P2X7, as P2X4 inhibition with PSB-12062 did not alter expression of IL-8 or E-selectin (see ). eNOS expression was unaffected by P2X7 inhibition (data not shown), while NFATc1 (but not other NFAT transcripts) was downregulated in endothelial cells exposed to atheroprone flow and this expression was further attenuated in the presence of the P2X7 antagonist (data not shown).

    Techniques: 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.

    Journal: Cardiovascular Research

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

    doi: 10.1093/cvr/cvx213

    Figure Lengend 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.

    Article Snippet: These effects were specific to P2X7, as P2X4 inhibition with PSB-12062 did not alter expression of IL-8 or E-selectin (see ). eNOS expression was unaffected by P2X7 inhibition (data not shown), while NFATc1 (but not other NFAT transcripts) was downregulated in endothelial cells exposed to atheroprone flow and this expression was further attenuated in the presence of the P2X7 antagonist (data not shown).

    Techniques: 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.

    Journal: Cardiovascular Research

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

    doi: 10.1093/cvr/cvx213

    Figure Lengend 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.

    Article Snippet: These effects were specific to P2X7, as P2X4 inhibition with PSB-12062 did not alter expression of IL-8 or E-selectin (see ). eNOS expression was unaffected by P2X7 inhibition (data not shown), while NFATc1 (but not other NFAT transcripts) was downregulated in endothelial cells exposed to atheroprone flow and this expression was further attenuated in the presence of the P2X7 antagonist (data not shown).

    Techniques: 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 =

    Journal: Cardiovascular Research

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

    doi: 10.1093/cvr/cvx213

    Figure Lengend 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 =

    Article Snippet: These effects were specific to P2X7, as P2X4 inhibition with PSB-12062 did not alter expression of IL-8 or E-selectin (see ). eNOS expression was unaffected by P2X7 inhibition (data not shown), while NFATc1 (but not other NFAT transcripts) was downregulated in endothelial cells exposed to atheroprone flow and this expression was further attenuated in the presence of the P2X7 antagonist (data not shown).

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