p2x 7 apr 008  (Alomone Labs)


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    Alomone Labs p2x 7 apr 008
    (A) Immortalized GEC that had been depleted of <t>P2X</t> 4 or <t>P2X</t> <t>7</t> using lentiviral particles were treated with 100 µM or 3 mM ATP for 3 hours, and supernatants were collected for caspase-1 activity measurement. Caspase-1 activity was measured by ELISA as described in . (B) Total proteins isolated from GEC were subjected to immunoprecipitation (IP) by a polyclonal anti-P2X 4 antibody or Dynabeads as a control. Precipitates or total protein extract (as input) were resolved on SDS-PAGE and analyzed on immunoblots with anti-P2X 7 (top), anti-pannexin-1 (middle), or anti-P2X 4 (bottom) antibodies.
    P2x 7 Apr 008, 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
    https://www.bioz.com/result/p2x 7 apr 008/product/Alomone Labs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p2x 7 apr 008 - by Bioz Stars, 2023-02
    95/100 stars

    Images

    1) Product Images from "P2X 4 Assembles with P2X 7 and Pannexin-1 in Gingival Epithelial Cells and Modulates ATP-induced Reactive Oxygen Species Production and Inflammasome Activation"

    Article Title: P2X 4 Assembles with P2X 7 and Pannexin-1 in Gingival Epithelial Cells and Modulates ATP-induced Reactive Oxygen Species Production and Inflammasome Activation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0070210

    (A) Immortalized GEC that had been depleted of P2X 4 or P2X 7 using lentiviral particles were treated with 100 µM or 3 mM ATP for 3 hours, and supernatants were collected for caspase-1 activity measurement. Caspase-1 activity was measured by ELISA as described in . (B) Total proteins isolated from GEC were subjected to immunoprecipitation (IP) by a polyclonal anti-P2X 4 antibody or Dynabeads as a control. Precipitates or total protein extract (as input) were resolved on SDS-PAGE and analyzed on immunoblots with anti-P2X 7 (top), anti-pannexin-1 (middle), or anti-P2X 4 (bottom) antibodies.
    Figure Legend Snippet: (A) Immortalized GEC that had been depleted of P2X 4 or P2X 7 using lentiviral particles were treated with 100 µM or 3 mM ATP for 3 hours, and supernatants were collected for caspase-1 activity measurement. Caspase-1 activity was measured by ELISA as described in . (B) Total proteins isolated from GEC were subjected to immunoprecipitation (IP) by a polyclonal anti-P2X 4 antibody or Dynabeads as a control. Precipitates or total protein extract (as input) were resolved on SDS-PAGE and analyzed on immunoblots with anti-P2X 7 (top), anti-pannexin-1 (middle), or anti-P2X 4 (bottom) antibodies.

    Techniques Used: Activity Assay, Enzyme-linked Immunosorbent Assay, Isolation, Immunoprecipitation, SDS Page, Western Blot

    Primary GEC (C and D) and immortalized GEC (A and B) were infected with or without P. gingivalis ( P.g. ) at an M.O.I. of 100 for 6 hours, followed by treatment with different pharmaceutical agents. Infected cells were treated with 100 µM ATP, 3 mM ATP, 3 mM ADP, 3 mM AMP, or 3 mM UTP individually for 1 hour (A and C). Alternatively, infected cells were pre-treated with 50 µM 5-BDBD for 15 minutes, 100 µM PPADS for 15 minutes, 100 µM oxATP for 30 minutes, or 1 mM probenecid for 10 minutes, followed by treatment with 3 mM ATP for 1 hour (B and D). The supernatants were collected and subjected to ELISA to measure IL-1β secretion. (E) Primary GEC were transfected with siRNA sequences against P2X 4 or P2X 7 for one day, and mRNA levels were detected by qPCR. (F) Primary GEC depleted of P2X 4 or P2X 7 were infected with P. gingivalis ( P.g. ) and treated with probenecid and 3 mM ATP as shown in (B). IL-1β secretion in the supernatants was analyzed by ELISA. The values showed averages and SD from duplicate samples, which were obtained from three separate experiments.
    Figure Legend Snippet: Primary GEC (C and D) and immortalized GEC (A and B) were infected with or without P. gingivalis ( P.g. ) at an M.O.I. of 100 for 6 hours, followed by treatment with different pharmaceutical agents. Infected cells were treated with 100 µM ATP, 3 mM ATP, 3 mM ADP, 3 mM AMP, or 3 mM UTP individually for 1 hour (A and C). Alternatively, infected cells were pre-treated with 50 µM 5-BDBD for 15 minutes, 100 µM PPADS for 15 minutes, 100 µM oxATP for 30 minutes, or 1 mM probenecid for 10 minutes, followed by treatment with 3 mM ATP for 1 hour (B and D). The supernatants were collected and subjected to ELISA to measure IL-1β secretion. (E) Primary GEC were transfected with siRNA sequences against P2X 4 or P2X 7 for one day, and mRNA levels were detected by qPCR. (F) Primary GEC depleted of P2X 4 or P2X 7 were infected with P. gingivalis ( P.g. ) and treated with probenecid and 3 mM ATP as shown in (B). IL-1β secretion in the supernatants was analyzed by ELISA. The values showed averages and SD from duplicate samples, which were obtained from three separate experiments.

    Techniques Used: Infection, Enzyme-linked Immunosorbent Assay, Transfection

    Model showing the role of P2X 4 and P2X 7 in ROS production and inflammasome activation in GEC stimulated with extracellular ATP.
    Figure Legend Snippet: Model showing the role of P2X 4 and P2X 7 in ROS production and inflammasome activation in GEC stimulated with extracellular ATP.

    Techniques Used: Activation Assay

    p2x 7 apr 008  (Alomone Labs)


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    Alomone Labs p2x 7 apr 008
    (A) Immortalized GEC that had been depleted of <t>P2X</t> 4 or <t>P2X</t> <t>7</t> using lentiviral particles were treated with 100 µM or 3 mM ATP for 3 hours, and supernatants were collected for caspase-1 activity measurement. Caspase-1 activity was measured by ELISA as described in . (B) Total proteins isolated from GEC were subjected to immunoprecipitation (IP) by a polyclonal anti-P2X 4 antibody or Dynabeads as a control. Precipitates or total protein extract (as input) were resolved on SDS-PAGE and analyzed on immunoblots with anti-P2X 7 (top), anti-pannexin-1 (middle), or anti-P2X 4 (bottom) antibodies.
    P2x 7 Apr 008, 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
    https://www.bioz.com/result/p2x 7 apr 008/product/Alomone Labs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p2x 7 apr 008 - by Bioz Stars, 2023-02
    95/100 stars

    Images

    1) Product Images from "P2X 4 Assembles with P2X 7 and Pannexin-1 in Gingival Epithelial Cells and Modulates ATP-induced Reactive Oxygen Species Production and Inflammasome Activation"

    Article Title: P2X 4 Assembles with P2X 7 and Pannexin-1 in Gingival Epithelial Cells and Modulates ATP-induced Reactive Oxygen Species Production and Inflammasome Activation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0070210

    (A) Immortalized GEC that had been depleted of P2X 4 or P2X 7 using lentiviral particles were treated with 100 µM or 3 mM ATP for 3 hours, and supernatants were collected for caspase-1 activity measurement. Caspase-1 activity was measured by ELISA as described in . (B) Total proteins isolated from GEC were subjected to immunoprecipitation (IP) by a polyclonal anti-P2X 4 antibody or Dynabeads as a control. Precipitates or total protein extract (as input) were resolved on SDS-PAGE and analyzed on immunoblots with anti-P2X 7 (top), anti-pannexin-1 (middle), or anti-P2X 4 (bottom) antibodies.
    Figure Legend Snippet: (A) Immortalized GEC that had been depleted of P2X 4 or P2X 7 using lentiviral particles were treated with 100 µM or 3 mM ATP for 3 hours, and supernatants were collected for caspase-1 activity measurement. Caspase-1 activity was measured by ELISA as described in . (B) Total proteins isolated from GEC were subjected to immunoprecipitation (IP) by a polyclonal anti-P2X 4 antibody or Dynabeads as a control. Precipitates or total protein extract (as input) were resolved on SDS-PAGE and analyzed on immunoblots with anti-P2X 7 (top), anti-pannexin-1 (middle), or anti-P2X 4 (bottom) antibodies.

    Techniques Used: Activity Assay, Enzyme-linked Immunosorbent Assay, Isolation, Immunoprecipitation, SDS Page, Western Blot

    Primary GEC (C and D) and immortalized GEC (A and B) were infected with or without P. gingivalis ( P.g. ) at an M.O.I. of 100 for 6 hours, followed by treatment with different pharmaceutical agents. Infected cells were treated with 100 µM ATP, 3 mM ATP, 3 mM ADP, 3 mM AMP, or 3 mM UTP individually for 1 hour (A and C). Alternatively, infected cells were pre-treated with 50 µM 5-BDBD for 15 minutes, 100 µM PPADS for 15 minutes, 100 µM oxATP for 30 minutes, or 1 mM probenecid for 10 minutes, followed by treatment with 3 mM ATP for 1 hour (B and D). The supernatants were collected and subjected to ELISA to measure IL-1β secretion. (E) Primary GEC were transfected with siRNA sequences against P2X 4 or P2X 7 for one day, and mRNA levels were detected by qPCR. (F) Primary GEC depleted of P2X 4 or P2X 7 were infected with P. gingivalis ( P.g. ) and treated with probenecid and 3 mM ATP as shown in (B). IL-1β secretion in the supernatants was analyzed by ELISA. The values showed averages and SD from duplicate samples, which were obtained from three separate experiments.
    Figure Legend Snippet: Primary GEC (C and D) and immortalized GEC (A and B) were infected with or without P. gingivalis ( P.g. ) at an M.O.I. of 100 for 6 hours, followed by treatment with different pharmaceutical agents. Infected cells were treated with 100 µM ATP, 3 mM ATP, 3 mM ADP, 3 mM AMP, or 3 mM UTP individually for 1 hour (A and C). Alternatively, infected cells were pre-treated with 50 µM 5-BDBD for 15 minutes, 100 µM PPADS for 15 minutes, 100 µM oxATP for 30 minutes, or 1 mM probenecid for 10 minutes, followed by treatment with 3 mM ATP for 1 hour (B and D). The supernatants were collected and subjected to ELISA to measure IL-1β secretion. (E) Primary GEC were transfected with siRNA sequences against P2X 4 or P2X 7 for one day, and mRNA levels were detected by qPCR. (F) Primary GEC depleted of P2X 4 or P2X 7 were infected with P. gingivalis ( P.g. ) and treated with probenecid and 3 mM ATP as shown in (B). IL-1β secretion in the supernatants was analyzed by ELISA. The values showed averages and SD from duplicate samples, which were obtained from three separate experiments.

    Techniques Used: Infection, Enzyme-linked Immunosorbent Assay, Transfection

    Model showing the role of P2X 4 and P2X 7 in ROS production and inflammasome activation in GEC stimulated with extracellular ATP.
    Figure Legend Snippet: Model showing the role of P2X 4 and P2X 7 in ROS production and inflammasome activation in GEC stimulated with extracellular ATP.

    Techniques Used: Activation Assay

    anti purinergic p2x 7 r antibody  (Alomone Labs)


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    Alomone Labs anti purinergic p2x 7 r antibody
    (A). Astrocytes were treated with PACAP (20 ng/ml), or indirectly co-cultured with hMSCs in the absence and presence of PACAP (20 ng/ml) for 48 hours. The cultures were subjected to Q-PCR for measurement of the mRNA levels of GLT-1 and GLAST. Data are presented as mean ± SEM and expressed as the ratio of GLT-1 (GLAST) mRNA levels compared to control. * p <0.05 versus control. (B). Astrocytes were indirectly co-cultured with hMSCs in the absence or presence of PACAP for 48 hours. Membrane proteins were then extracted for western blotting using anti-GLT-1 antibody. The same blot was reprobed with <t>anti-P2X</t> 7 R antibody. <t>P2X</t> <t>7</t> <t>R</t> levels are presented as a loading control. Relative intensity of GLT-1 levels normalized to P2X 7 R was measured. Data are presented as mean ± SEM and expressed as a percentage of GLT-1 levels in the group with combinatorial treatment compared to that detected in the group co-cultured only with hMSCs. * p <0.05 versus the group only co-cultured with hMSCs. (C). Astrocytes were infected by lentivirus carrying lenti-GLT-1-GFP, and then co-cultured with hMSCs in the absence or presence of PACAP. Strong punctate fluorescence spots (arrowheads) were observed in the processes of astrocytes co-cultured with hMSCs in the presence of PACAP, when compared to that seen in astrocytes co-cultured only with hMSCs (arrows). Scale bar, 50 µm. (D). Astrocytes were treated with PACAP, or co-cultured with hMSCs in the absence or presence of PACAP. After 48 or 72 hours, the cultures were subjected to [ 3 H]-L-glutamate uptake analysis as described in . Data are presented as mean ± SEM and expressed as a ratio of the glutamate uptake in each treated group compared to that detected in the control group. * p <0.05 versus control.
    Anti Purinergic P2x 7 R Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti purinergic p2x 7 r antibody/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti purinergic p2x 7 r antibody - by Bioz Stars, 2023-02
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    Images

    1) Product Images from "Effects of Combinatorial Treatment with Pituitary Adenylate Cyclase Activating Peptide and Human Mesenchymal Stem Cells on Spinal Cord Tissue Repair"

    Article Title: Effects of Combinatorial Treatment with Pituitary Adenylate Cyclase Activating Peptide and Human Mesenchymal Stem Cells on Spinal Cord Tissue Repair

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0015299

    (A). Astrocytes were treated with PACAP (20 ng/ml), or indirectly co-cultured with hMSCs in the absence and presence of PACAP (20 ng/ml) for 48 hours. The cultures were subjected to Q-PCR for measurement of the mRNA levels of GLT-1 and GLAST. Data are presented as mean ± SEM and expressed as the ratio of GLT-1 (GLAST) mRNA levels compared to control. * p <0.05 versus control. (B). Astrocytes were indirectly co-cultured with hMSCs in the absence or presence of PACAP for 48 hours. Membrane proteins were then extracted for western blotting using anti-GLT-1 antibody. The same blot was reprobed with anti-P2X 7 R antibody. P2X 7 R levels are presented as a loading control. Relative intensity of GLT-1 levels normalized to P2X 7 R was measured. Data are presented as mean ± SEM and expressed as a percentage of GLT-1 levels in the group with combinatorial treatment compared to that detected in the group co-cultured only with hMSCs. * p <0.05 versus the group only co-cultured with hMSCs. (C). Astrocytes were infected by lentivirus carrying lenti-GLT-1-GFP, and then co-cultured with hMSCs in the absence or presence of PACAP. Strong punctate fluorescence spots (arrowheads) were observed in the processes of astrocytes co-cultured with hMSCs in the presence of PACAP, when compared to that seen in astrocytes co-cultured only with hMSCs (arrows). Scale bar, 50 µm. (D). Astrocytes were treated with PACAP, or co-cultured with hMSCs in the absence or presence of PACAP. After 48 or 72 hours, the cultures were subjected to [ 3 H]-L-glutamate uptake analysis as described in . Data are presented as mean ± SEM and expressed as a ratio of the glutamate uptake in each treated group compared to that detected in the control group. * p <0.05 versus control.
    Figure Legend Snippet: (A). Astrocytes were treated with PACAP (20 ng/ml), or indirectly co-cultured with hMSCs in the absence and presence of PACAP (20 ng/ml) for 48 hours. The cultures were subjected to Q-PCR for measurement of the mRNA levels of GLT-1 and GLAST. Data are presented as mean ± SEM and expressed as the ratio of GLT-1 (GLAST) mRNA levels compared to control. * p <0.05 versus control. (B). Astrocytes were indirectly co-cultured with hMSCs in the absence or presence of PACAP for 48 hours. Membrane proteins were then extracted for western blotting using anti-GLT-1 antibody. The same blot was reprobed with anti-P2X 7 R antibody. P2X 7 R levels are presented as a loading control. Relative intensity of GLT-1 levels normalized to P2X 7 R was measured. Data are presented as mean ± SEM and expressed as a percentage of GLT-1 levels in the group with combinatorial treatment compared to that detected in the group co-cultured only with hMSCs. * p <0.05 versus the group only co-cultured with hMSCs. (C). Astrocytes were infected by lentivirus carrying lenti-GLT-1-GFP, and then co-cultured with hMSCs in the absence or presence of PACAP. Strong punctate fluorescence spots (arrowheads) were observed in the processes of astrocytes co-cultured with hMSCs in the presence of PACAP, when compared to that seen in astrocytes co-cultured only with hMSCs (arrows). Scale bar, 50 µm. (D). Astrocytes were treated with PACAP, or co-cultured with hMSCs in the absence or presence of PACAP. After 48 or 72 hours, the cultures were subjected to [ 3 H]-L-glutamate uptake analysis as described in . Data are presented as mean ± SEM and expressed as a ratio of the glutamate uptake in each treated group compared to that detected in the control group. * p <0.05 versus control.

    Techniques Used: Cell Culture, Western Blot, Infection, Fluorescence

    affinity purified rabbit anti p2x7r antibodies  (Alomone Labs)


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    Alomone Labs affinity purified rabbit anti p2x7r antibodies
    Spleen cells from 3 to 4-mo-old MRL +/+ ( solid line ), MRL/ lpr ( dashed line ) and P2X7-deficient B6 <t>P2X7R−/−</t> ( dotted line ) mice (n = 3 mice/group) were treated for 45 min at 37°C with doses of ATP ranging from 100 to 5000 µM. Spleen cells were then triple-stained with anti-CD90, anti-CD19 and anti-CD62L mAb to assess by flow cytometry: (A) the percentage of CD62L-expressing CD19 – CD90 + T cells and (B) MFI of CD62L on CD19 – CD90 + T cells. Results are expressed as the mean percentage of initial expression ± SE.
    Affinity Purified Rabbit Anti P2x7r Antibodies, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/affinity purified rabbit anti p2x7r antibodies/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    affinity purified rabbit anti p2x7r antibodies - by Bioz Stars, 2023-02
    94/100 stars

    Images

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

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

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0052161

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

    Techniques Used: Staining, Flow Cytometry, Expressing

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

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

    p2x7 receptors  (Alomone Labs)


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    Alomone Labs p2x7 receptors
    Intracellular Ca 2+ measurement (using Fura-2-AM) in macrophages subjected to treatment with the <t>P2X7</t> agonist BzATP and/or with the antagonist A740003. (a) Changes in intracellular Ca 2+ levels in macrophages from control (black line) and S. mansoni -infected (red line) mice. Inset: representative images from macrophages (control) before (baseline) and after treatment with 100 μ M BzATP. (b) Changes in intracellular Ca 2+ levels ([Ca 2+ ] i ) in macrophages from control (white bars) or S. mansoni- infected (black bars) mice. Three plates were used for each condition/animal, 10 cells/plate were chosen randomly for imaging, and cells were obtained from three animals. Data were expressed as mean and SEM. * P < 0.05; *** P < 0.001 (one-way ANOVA followed by post hoc Newman-Keuls test).
    P2x7 Receptors, 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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    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p2x7 receptors - by Bioz Stars, 2023-02
    95/100 stars

    Images

    1) Product Images from "Macrophage P2X7 Receptor Function Is Reduced during Schistosomiasis: Putative Role of TGF- β 1"

    Article Title: Macrophage P2X7 Receptor Function Is Reduced during Schistosomiasis: Putative Role of TGF- β 1

    Journal: Mediators of Inflammation

    doi: 10.1155/2014/134974

    Intracellular Ca 2+ measurement (using Fura-2-AM) in macrophages subjected to treatment with the P2X7 agonist BzATP and/or with the antagonist A740003. (a) Changes in intracellular Ca 2+ levels in macrophages from control (black line) and S. mansoni -infected (red line) mice. Inset: representative images from macrophages (control) before (baseline) and after treatment with 100 μ M BzATP. (b) Changes in intracellular Ca 2+ levels ([Ca 2+ ] i ) in macrophages from control (white bars) or S. mansoni- infected (black bars) mice. Three plates were used for each condition/animal, 10 cells/plate were chosen randomly for imaging, and cells were obtained from three animals. Data were expressed as mean and SEM. * P < 0.05; *** P < 0.001 (one-way ANOVA followed by post hoc Newman-Keuls test).
    Figure Legend Snippet: Intracellular Ca 2+ measurement (using Fura-2-AM) in macrophages subjected to treatment with the P2X7 agonist BzATP and/or with the antagonist A740003. (a) Changes in intracellular Ca 2+ levels in macrophages from control (black line) and S. mansoni -infected (red line) mice. Inset: representative images from macrophages (control) before (baseline) and after treatment with 100 μ M BzATP. (b) Changes in intracellular Ca 2+ levels ([Ca 2+ ] i ) in macrophages from control (white bars) or S. mansoni- infected (black bars) mice. Three plates were used for each condition/animal, 10 cells/plate were chosen randomly for imaging, and cells were obtained from three animals. Data were expressed as mean and SEM. * P < 0.05; *** P < 0.001 (one-way ANOVA followed by post hoc Newman-Keuls test).

    Techniques Used: Infection, Imaging

    P2X7 receptors expression in peritoneal macrophages. (a) Macrophages from uninfected (white bar) and S. mansoni -infected (black bar) mice express similar levels of P2X7 protein. n = 9 replicates from three different animals, for each group. The images above are representative Western blots (U = uninfected and I = infected). (b) P2X7 receptors expression in peritoneal macrophages from control mice (white bar, untreated) or TGF- β 1-treated cells (5 ng/mL, 24 h; black bar) are also similar. The images above are representative Western blots (U = uninfected; TGF- β 1= TGF- β 1-treated macrophages). Lysates from P2X7 KO macrophages were used as negative controls for P2X7 protein expression. Cells lysates were obtained from three animals for each group. (c) Quantitative RT-PCR (qRT-PCR) analysis of P2X7 receptor mRNA levels in peritoneal macrophages. Similar levels of P2X7 mRNA were found in uninfected (white bar), S. mansoni -infected mice (black bar), and TGF- β 1-treated (5 ng/mL, 24 h; hatched bar) mouse macrophages. N = 3–5 samples per group (using different animals).
    Figure Legend Snippet: P2X7 receptors expression in peritoneal macrophages. (a) Macrophages from uninfected (white bar) and S. mansoni -infected (black bar) mice express similar levels of P2X7 protein. n = 9 replicates from three different animals, for each group. The images above are representative Western blots (U = uninfected and I = infected). (b) P2X7 receptors expression in peritoneal macrophages from control mice (white bar, untreated) or TGF- β 1-treated cells (5 ng/mL, 24 h; black bar) are also similar. The images above are representative Western blots (U = uninfected; TGF- β 1= TGF- β 1-treated macrophages). Lysates from P2X7 KO macrophages were used as negative controls for P2X7 protein expression. Cells lysates were obtained from three animals for each group. (c) Quantitative RT-PCR (qRT-PCR) analysis of P2X7 receptor mRNA levels in peritoneal macrophages. Similar levels of P2X7 mRNA were found in uninfected (white bar), S. mansoni -infected mice (black bar), and TGF- β 1-treated (5 ng/mL, 24 h; hatched bar) mouse macrophages. N = 3–5 samples per group (using different animals).

    Techniques Used: Expressing, Infection, Western Blot, Quantitative RT-PCR

    Confocal microscopy analysis of P2X7 receptor expression on the cell surface of macrophages. Cells were labelled with antibodies recognizing P2X7 receptors (green) and F4/80+ (red) as well as with DAPI (blue). (a) Representative images showing macrophages from uninfected (left column), S. mansoni -infected (middle column), and TGF- β 1-treated (right column; 5 ng/mL, for 24 h) mice. The bottom row shows orthogonal slices of the representative labelled cells, with red arrows marking P2X7 expression on the plasma membrane. (b) Quantitative analysis of confocal microscopy data. The mean fluorescence intensity, showing cell surface P2X7 receptor expression in peritoneal macrophages from uninfected mice (white bar), S. mansoni -infected (black bar), and TGF- β 1-treated (hatched bar; 5 ng/mL, for 24 h) mice, was obtained from pixels intensity using ImageJ software (see methods). Inset: negative controls (macrophages incubated with secondary antibodies only). n = 6 replicates from 3 animals for each condition (*** P < 0.001 versus control, P < 0.01 TGF- β 1 versus infected group; one-way ANOVA followed by post hoc Newman-Keuls test).
    Figure Legend Snippet: Confocal microscopy analysis of P2X7 receptor expression on the cell surface of macrophages. Cells were labelled with antibodies recognizing P2X7 receptors (green) and F4/80+ (red) as well as with DAPI (blue). (a) Representative images showing macrophages from uninfected (left column), S. mansoni -infected (middle column), and TGF- β 1-treated (right column; 5 ng/mL, for 24 h) mice. The bottom row shows orthogonal slices of the representative labelled cells, with red arrows marking P2X7 expression on the plasma membrane. (b) Quantitative analysis of confocal microscopy data. The mean fluorescence intensity, showing cell surface P2X7 receptor expression in peritoneal macrophages from uninfected mice (white bar), S. mansoni -infected (black bar), and TGF- β 1-treated (hatched bar; 5 ng/mL, for 24 h) mice, was obtained from pixels intensity using ImageJ software (see methods). Inset: negative controls (macrophages incubated with secondary antibodies only). n = 6 replicates from 3 animals for each condition (*** P < 0.001 versus control, P < 0.01 TGF- β 1 versus infected group; one-way ANOVA followed by post hoc Newman-Keuls test).

    Techniques Used: Confocal Microscopy, Expressing, Infection, Fluorescence, Software, Incubation

    Survival curve of C57BL/6 wild type or P2X7 KO infected with approximately 80 cercariae of S. mansoni . (P2X7 KO: n = 9 females and 7 males; C57BL/6 wild type: 4 females and 10 males). *** P < 0.0001 using the Mantel-Cox log-rank test.
    Figure Legend Snippet: Survival curve of C57BL/6 wild type or P2X7 KO infected with approximately 80 cercariae of S. mansoni . (P2X7 KO: n = 9 females and 7 males; C57BL/6 wild type: 4 females and 10 males). *** P < 0.0001 using the Mantel-Cox log-rank test.

    Techniques Used: Infection

    rabbit anti p2x7r monoclonal igg  (Alomone Labs)


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    Structured Review

    Alomone Labs rabbit anti p2x7r monoclonal igg
    The sequences of oligonucleotide primers.
    Rabbit Anti P2x7r Monoclonal Igg, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti p2x7r monoclonal igg/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti p2x7r monoclonal igg - by Bioz Stars, 2023-02
    94/100 stars

    Images

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

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

    Journal: BioMed Research International

    doi: 10.1155/2020/8143754

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

    Techniques Used:

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

    Techniques Used: Western Blot, Concentration Assay

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

    Techniques Used: Fluorescence, Labeling, Concentration Assay

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

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

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

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

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

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

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

    Techniques Used: Expressing, Inhibition

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

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

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

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

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

    Techniques Used: Inhibition

    rabbit anti p2x7r monoclonal igg  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
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  • 94

    Structured Review

    Alomone Labs rabbit anti p2x7r monoclonal igg
    The sequences of oligonucleotide primers.
    Rabbit Anti P2x7r Monoclonal Igg, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti p2x7r monoclonal igg/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti p2x7r monoclonal igg - by Bioz Stars, 2023-02
    94/100 stars

    Images

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

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

    Journal: BioMed Research International

    doi: 10.1155/2020/8143754

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

    Techniques Used:

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

    Techniques Used: Western Blot, Concentration Assay

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

    Techniques Used: Fluorescence, Labeling, Concentration Assay

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

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

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

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

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

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

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

    Techniques Used: Expressing, Inhibition

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

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

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

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

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

    Techniques Used: Inhibition

    anti p2x7 polyclonal rabbit antibody  (Alomone Labs)


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  • 95

    Structured Review

    Alomone Labs anti p2x7 polyclonal rabbit antibody
    Association of pannexin1 and <t>P2X7</t> in unwounded corneas and 2 hrs after injury in DiO and B6 control corneal epithelium. Proximity ligation assays were performed using antibodies to P2X7 and pannexin1. (a) Representative enface and orthogonal (ortho) images of B6 control and DiO corneas are shown. The green color displays puncta of association. Corneal epithelium was counter stained with rhodamine phalloidin (asterisk marks wound area). The numbers and letters mark the individual cells that were analyzed using CellProfiler near the wound and back from the wound. The boxes indicate the representative individual cells that are shown in (b) after PLA analysis with CellProfiler. Bar equals 20 microns (PLA enface and actin) or 15 microns (PLA ortho). (b) Analysis of overlapping puncta for each condition (unwounded, leading edge, back from leading edge) was performed and a box plot drawn using R. The mean puncta for each region are placed above the box. Mean ± SEM are plotted, and two-way ANOVA with Tukey's multiple comparison of means was performed. p < 0.05. N is a minimum of 3 independent experiments.
    Anti P2x7 Polyclonal Rabbit Antibody, 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
    https://www.bioz.com/result/anti p2x7 polyclonal rabbit antibody/product/Alomone Labs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti p2x7 polyclonal rabbit antibody - by Bioz Stars, 2023-02
    95/100 stars

    Images

    1) Product Images from "Pannexin1: Role as a Sensor to Injury Is Attenuated in Pretype 2 Corneal Diabetic Epithelium"

    Article Title: Pannexin1: Role as a Sensor to Injury Is Attenuated in Pretype 2 Corneal Diabetic Epithelium

    Journal: Analytical Cellular Pathology (Amsterdam)

    doi: 10.1155/2021/4793338

    Association of pannexin1 and P2X7 in unwounded corneas and 2 hrs after injury in DiO and B6 control corneal epithelium. Proximity ligation assays were performed using antibodies to P2X7 and pannexin1. (a) Representative enface and orthogonal (ortho) images of B6 control and DiO corneas are shown. The green color displays puncta of association. Corneal epithelium was counter stained with rhodamine phalloidin (asterisk marks wound area). The numbers and letters mark the individual cells that were analyzed using CellProfiler near the wound and back from the wound. The boxes indicate the representative individual cells that are shown in (b) after PLA analysis with CellProfiler. Bar equals 20 microns (PLA enface and actin) or 15 microns (PLA ortho). (b) Analysis of overlapping puncta for each condition (unwounded, leading edge, back from leading edge) was performed and a box plot drawn using R. The mean puncta for each region are placed above the box. Mean ± SEM are plotted, and two-way ANOVA with Tukey's multiple comparison of means was performed. p < 0.05. N is a minimum of 3 independent experiments.
    Figure Legend Snippet: Association of pannexin1 and P2X7 in unwounded corneas and 2 hrs after injury in DiO and B6 control corneal epithelium. Proximity ligation assays were performed using antibodies to P2X7 and pannexin1. (a) Representative enface and orthogonal (ortho) images of B6 control and DiO corneas are shown. The green color displays puncta of association. Corneal epithelium was counter stained with rhodamine phalloidin (asterisk marks wound area). The numbers and letters mark the individual cells that were analyzed using CellProfiler near the wound and back from the wound. The boxes indicate the representative individual cells that are shown in (b) after PLA analysis with CellProfiler. Bar equals 20 microns (PLA enface and actin) or 15 microns (PLA ortho). (b) Analysis of overlapping puncta for each condition (unwounded, leading edge, back from leading edge) was performed and a box plot drawn using R. The mean puncta for each region are placed above the box. Mean ± SEM are plotted, and two-way ANOVA with Tukey's multiple comparison of means was performed. p < 0.05. N is a minimum of 3 independent experiments.

    Techniques Used: Ligation, Staining

    p2x7 receptors  (Alomone Labs)


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    Alomone Labs p2x7 receptors
    Association of pannexin1 and <t>P2X7</t> in unwounded corneas and 2 hrs after injury in DiO and B6 control corneal epithelium. Proximity ligation assays were performed using antibodies to P2X7 and pannexin1. (a) Representative enface and orthogonal (ortho) images of B6 control and DiO corneas are shown. The green color displays puncta of association. Corneal epithelium was counter stained with rhodamine phalloidin (asterisk marks wound area). The numbers and letters mark the individual cells that were analyzed using CellProfiler near the wound and back from the wound. The boxes indicate the representative individual cells that are shown in (b) after PLA analysis with CellProfiler. Bar equals 20 microns (PLA enface and actin) or 15 microns (PLA ortho). (b) Analysis of overlapping puncta for each condition (unwounded, leading edge, back from leading edge) was performed and a box plot drawn using R. The mean puncta for each region are placed above the box. Mean ± SEM are plotted, and two-way ANOVA with Tukey's multiple comparison of means was performed. p < 0.05. N is a minimum of 3 independent experiments.
    P2x7 Receptors, 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
    https://www.bioz.com/result/p2x7 receptors/product/Alomone Labs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p2x7 receptors - by Bioz Stars, 2023-02
    95/100 stars

    Images

    1) Product Images from "Pannexin1: Role as a Sensor to Injury Is Attenuated in Pretype 2 Corneal Diabetic Epithelium"

    Article Title: Pannexin1: Role as a Sensor to Injury Is Attenuated in Pretype 2 Corneal Diabetic Epithelium

    Journal: Analytical Cellular Pathology (Amsterdam)

    doi: 10.1155/2021/4793338

    Association of pannexin1 and P2X7 in unwounded corneas and 2 hrs after injury in DiO and B6 control corneal epithelium. Proximity ligation assays were performed using antibodies to P2X7 and pannexin1. (a) Representative enface and orthogonal (ortho) images of B6 control and DiO corneas are shown. The green color displays puncta of association. Corneal epithelium was counter stained with rhodamine phalloidin (asterisk marks wound area). The numbers and letters mark the individual cells that were analyzed using CellProfiler near the wound and back from the wound. The boxes indicate the representative individual cells that are shown in (b) after PLA analysis with CellProfiler. Bar equals 20 microns (PLA enface and actin) or 15 microns (PLA ortho). (b) Analysis of overlapping puncta for each condition (unwounded, leading edge, back from leading edge) was performed and a box plot drawn using R. The mean puncta for each region are placed above the box. Mean ± SEM are plotted, and two-way ANOVA with Tukey's multiple comparison of means was performed. p < 0.05. N is a minimum of 3 independent experiments.
    Figure Legend Snippet: Association of pannexin1 and P2X7 in unwounded corneas and 2 hrs after injury in DiO and B6 control corneal epithelium. Proximity ligation assays were performed using antibodies to P2X7 and pannexin1. (a) Representative enface and orthogonal (ortho) images of B6 control and DiO corneas are shown. The green color displays puncta of association. Corneal epithelium was counter stained with rhodamine phalloidin (asterisk marks wound area). The numbers and letters mark the individual cells that were analyzed using CellProfiler near the wound and back from the wound. The boxes indicate the representative individual cells that are shown in (b) after PLA analysis with CellProfiler. Bar equals 20 microns (PLA enface and actin) or 15 microns (PLA ortho). (b) Analysis of overlapping puncta for each condition (unwounded, leading edge, back from leading edge) was performed and a box plot drawn using R. The mean puncta for each region are placed above the box. Mean ± SEM are plotted, and two-way ANOVA with Tukey's multiple comparison of means was performed. p < 0.05. N is a minimum of 3 independent experiments.

    Techniques Used: Ligation, Staining

    p2x7 r peptide  (Alomone Labs)


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    Structured Review

    Alomone Labs p2x7 r peptide
    Oligonucleotide Primer List.
    P2x7 R Peptide, 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
    https://www.bioz.com/result/p2x7 r peptide/product/Alomone Labs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p2x7 r peptide - by Bioz Stars, 2023-02
    95/100 stars

    Images

    1) Product Images from "Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse"

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0029990

    Oligonucleotide Primer List.
    Figure Legend Snippet: Oligonucleotide Primer List.

    Techniques Used:

    Antibody List.
    Figure Legend Snippet: Antibody List.

    Techniques Used: Binding Assay, Transduction

    To assess whether P2X7-R splice variants other than the full length, variant 1 are expressed in the WT and P2X7-KO mouse retina and also whether read through of variant 1 C-terminal mRNA occurs in the P2X7-KO mouse retina, PCR for (A) the central region of the P2X7-R which is common to variants 1, 2 and 3; (B) the C-terminus from within the start of the disruption in the P2X7-KO to the end of the mRNA (variant 1 only, primer pair A); and (C) the C-terminus from after the insertion deletion in the P2X7-KO mouse to the end of the receptor (variant 1 only, primer pair B) was completed. P2X7-R variant 1 C-terminal mRNA was absent (B–C) in the P2X7-KO mouse but N-terminal mRNA variants (likely variant 2 or 3) were still expressed (A). MW, molecular weight marker and the bright bar is 300 bp. (D–E) Protein analysis of P2X7-R splice variant expression was completed using Western blots of mouse retina (D) and cortex (E). An N-terminal specific P2X7-R antibody detected the protein transcript of variant 1 mRNA in WT samples 1 and 2 but not P2X7-KO samples 3 and 4 (68.41 kDa) of both retina and cortex. Protein for variant 2/3 (50.69 kDa/49.38 kDa) and 4 (17.2 kDa) was expressed in both WT and P2X7-KO retina and cortex. A monoclonal antibody against GAPDH was applied to the same blots and indicates that similar amounts of protein were loaded for WT and P2X7-KO samples.
    Figure Legend Snippet: To assess whether P2X7-R splice variants other than the full length, variant 1 are expressed in the WT and P2X7-KO mouse retina and also whether read through of variant 1 C-terminal mRNA occurs in the P2X7-KO mouse retina, PCR for (A) the central region of the P2X7-R which is common to variants 1, 2 and 3; (B) the C-terminus from within the start of the disruption in the P2X7-KO to the end of the mRNA (variant 1 only, primer pair A); and (C) the C-terminus from after the insertion deletion in the P2X7-KO mouse to the end of the receptor (variant 1 only, primer pair B) was completed. P2X7-R variant 1 C-terminal mRNA was absent (B–C) in the P2X7-KO mouse but N-terminal mRNA variants (likely variant 2 or 3) were still expressed (A). MW, molecular weight marker and the bright bar is 300 bp. (D–E) Protein analysis of P2X7-R splice variant expression was completed using Western blots of mouse retina (D) and cortex (E). An N-terminal specific P2X7-R antibody detected the protein transcript of variant 1 mRNA in WT samples 1 and 2 but not P2X7-KO samples 3 and 4 (68.41 kDa) of both retina and cortex. Protein for variant 2/3 (50.69 kDa/49.38 kDa) and 4 (17.2 kDa) was expressed in both WT and P2X7-KO retina and cortex. A monoclonal antibody against GAPDH was applied to the same blots and indicates that similar amounts of protein were loaded for WT and P2X7-KO samples.

    Techniques Used: Variant Assay, Molecular Weight, Marker, Expressing, Western Blot

    Mice were dark adapted overnight and mixed (rod and cone) ERG responses were recorded in response to full field flashes from intensity −4.2 to 2.1 log cd.s/m 2 . (A–B) Representative mixed ERG waveforms for (A) WT and (B) P2X7-KO mice are presented. (C) There was no change in the amplitude of the rod photoreceptor derived component, a-wave, as a function of intensity. (D) The amplitude of the post-photoreceptor response, the b-wave was larger in P2X7-KO mice than in WT mice across almost all intensities tested. * indicates p<0.05, a significant difference between WT and P2X7-KO.
    Figure Legend Snippet: Mice were dark adapted overnight and mixed (rod and cone) ERG responses were recorded in response to full field flashes from intensity −4.2 to 2.1 log cd.s/m 2 . (A–B) Representative mixed ERG waveforms for (A) WT and (B) P2X7-KO mice are presented. (C) There was no change in the amplitude of the rod photoreceptor derived component, a-wave, as a function of intensity. (D) The amplitude of the post-photoreceptor response, the b-wave was larger in P2X7-KO mice than in WT mice across almost all intensities tested. * indicates p<0.05, a significant difference between WT and P2X7-KO.

    Techniques Used: Derivative Assay

    (A) Representative waveforms of the rod ERG recorded from WT (black) and P2X7-KO mice (grey) in response to a 2.1 log cd.s/m 2 intensity flash. The photoreceptor derived, rod PIII (modelled a-wave) was not altered in either (B) amplitude or (C) sensitivity in the P2X7-KO mouse when compared with the WT response. The post-photoreceptor, rod PII (modelled b-wave) was found to be (D) larger in amplitude and (E) faster to reach a maximum response. (F) The amplitude of the third of the inner retinal derived oscillatory potentials was significantly enhanced in the P2X7-KO mouse. * indicates p<0.05, a significant difference between WT and P2X7-KO.
    Figure Legend Snippet: (A) Representative waveforms of the rod ERG recorded from WT (black) and P2X7-KO mice (grey) in response to a 2.1 log cd.s/m 2 intensity flash. The photoreceptor derived, rod PIII (modelled a-wave) was not altered in either (B) amplitude or (C) sensitivity in the P2X7-KO mouse when compared with the WT response. The post-photoreceptor, rod PII (modelled b-wave) was found to be (D) larger in amplitude and (E) faster to reach a maximum response. (F) The amplitude of the third of the inner retinal derived oscillatory potentials was significantly enhanced in the P2X7-KO mouse. * indicates p<0.05, a significant difference between WT and P2X7-KO.

    Techniques Used: Derivative Assay

    (A) Representative waveforms of the cone ERG recorded from WT (black) and P2X7-KO mice (grey) in response to a 2.1 log cd.s/m 2 intensity flash. The post-photoreceptor, cone PII (modelled b-wave) was found to be larger in (B) amplitude but not faster to reach a maximum response (C) in the P2X7-KO mouse when compared with the WT response. (D) The photopic visual acuity, assessed using the optokinetic reflex to determine the spatial frequency threshold, was higher in the P2X7-KO mouse than in the WT. * indicates p<0.05, a significant difference between WT and P2X7-KO.
    Figure Legend Snippet: (A) Representative waveforms of the cone ERG recorded from WT (black) and P2X7-KO mice (grey) in response to a 2.1 log cd.s/m 2 intensity flash. The post-photoreceptor, cone PII (modelled b-wave) was found to be larger in (B) amplitude but not faster to reach a maximum response (C) in the P2X7-KO mouse when compared with the WT response. (D) The photopic visual acuity, assessed using the optokinetic reflex to determine the spatial frequency threshold, was higher in the P2X7-KO mouse than in the WT. * indicates p<0.05, a significant difference between WT and P2X7-KO.

    Techniques Used:

    Transverse sections of paraffin embedded retina from (A) WT and (B) P2X7-KO mice were stained with hematoxylin and eosin for comparison. The gross retinal morphology was similar in both mice. Scale bar, 20 microns.
    Figure Legend Snippet: Transverse sections of paraffin embedded retina from (A) WT and (B) P2X7-KO mice were stained with hematoxylin and eosin for comparison. The gross retinal morphology was similar in both mice. Scale bar, 20 microns.

    Techniques Used: Staining

    (A) Immunohistochemistry on a transverse retinal section from a WT mouse showing P2X7-R immunoreactivity in the outer plexiform layer (OPL), inner nuclear layer (INL), and ganglion cell layer (GCL). (C) The retina does not label with the P2X7-R antibody following incubation with P2X7-R specific peptide. Scale bar, 20 microns.
    Figure Legend Snippet: (A) Immunohistochemistry on a transverse retinal section from a WT mouse showing P2X7-R immunoreactivity in the outer plexiform layer (OPL), inner nuclear layer (INL), and ganglion cell layer (GCL). (C) The retina does not label with the P2X7-R antibody following incubation with P2X7-R specific peptide. Scale bar, 20 microns.

    Techniques Used: Immunohistochemistry, Incubation

    (A) P2X7-R IR (green) colocalises with horizontal cells labelled with Calbindin (red) and their terminals. (B) P2X7-R IR (green) colocalises with rod bipolar cells labelled with PKC (red). (C) Inset of rod bipolar cell terminals. (D) P2X7-R IR (green) puncta are discretely expressed within photoreceptor terminals labelled with PSD-95 (red). (E) Cone bipolar cells (Type 2 and 6) labelled with a ZNP-1 antibody (green) colocalise with P2X7-R IR (red) including both the cone bipolar cell dendrites and terminals (F). P2X7-R IR (green) colocalises with amacrine cells labelled with calretinin (G) and in particular GABAergic amacrine cells (H). (I) Ganglion cells labelled with an antibody against GFP in the Thy1-HYFP mouse (pseudo-coloured red) were P2X7-R IR (pseudo-coloured green). Scale bar, 10 microns.
    Figure Legend Snippet: (A) P2X7-R IR (green) colocalises with horizontal cells labelled with Calbindin (red) and their terminals. (B) P2X7-R IR (green) colocalises with rod bipolar cells labelled with PKC (red). (C) Inset of rod bipolar cell terminals. (D) P2X7-R IR (green) puncta are discretely expressed within photoreceptor terminals labelled with PSD-95 (red). (E) Cone bipolar cells (Type 2 and 6) labelled with a ZNP-1 antibody (green) colocalise with P2X7-R IR (red) including both the cone bipolar cell dendrites and terminals (F). P2X7-R IR (green) colocalises with amacrine cells labelled with calretinin (G) and in particular GABAergic amacrine cells (H). (I) Ganglion cells labelled with an antibody against GFP in the Thy1-HYFP mouse (pseudo-coloured red) were P2X7-R IR (pseudo-coloured green). Scale bar, 10 microns.

    Techniques Used:

    (A) P2X7-R IR (red) is associated with but does not colocalise with ribbon synapses (ribeye, green) on bipolar cell terminals (VGLUT, blue). (B) Magnified inset from A, arrows indicate ribbon synapses associated with P2X7-R IR puncta. (C) P2X7-R IR (red) colocalises with conventional synapses (bassoon, green) distinct from bipolar cell terminals (VGLUT, blue). (D) Magnified inset from C, arrows indicate conventional synapses that colocalise with P2X7-R IR puncta. Co-localisation analysis indicates that P2X7-R IR puncta do not co-localise with ribbon synapses (E) but do show significant colocalisation with conventional synapses (F) in the IPL. Scale bars are 5 microns for A & C and 2 microns for B & D.
    Figure Legend Snippet: (A) P2X7-R IR (red) is associated with but does not colocalise with ribbon synapses (ribeye, green) on bipolar cell terminals (VGLUT, blue). (B) Magnified inset from A, arrows indicate ribbon synapses associated with P2X7-R IR puncta. (C) P2X7-R IR (red) colocalises with conventional synapses (bassoon, green) distinct from bipolar cell terminals (VGLUT, blue). (D) Magnified inset from C, arrows indicate conventional synapses that colocalise with P2X7-R IR puncta. Co-localisation analysis indicates that P2X7-R IR puncta do not co-localise with ribbon synapses (E) but do show significant colocalisation with conventional synapses (F) in the IPL. Scale bars are 5 microns for A & C and 2 microns for B & D.

    Techniques Used:

    Ribbons are indicated by arrows and labelled amacine cells are indicated by astericks. (A), (B) & (C) P2X7-R immunoreactivity labels amacrine cells adjacent to rod bipolar cell ribbon synapses in the IPL. (D) A conventional amacrine cell synapse is labelled for P2X7 immunoreactivity in close proximity to a rod bipolar cell terminal. Scale bars are 200 nm for A & B and 500 nm for C & D.
    Figure Legend Snippet: Ribbons are indicated by arrows and labelled amacine cells are indicated by astericks. (A), (B) & (C) P2X7-R immunoreactivity labels amacrine cells adjacent to rod bipolar cell ribbon synapses in the IPL. (D) A conventional amacrine cell synapse is labelled for P2X7 immunoreactivity in close proximity to a rod bipolar cell terminal. Scale bars are 200 nm for A & B and 500 nm for C & D.

    Techniques Used:

    (A–B) Müller cells, labelled with an antibody against glutamine synthetise (red) were found to be closely associated with P2X7-R-IR puncta in the IPL (green), but not co-localised in the IPL. (C) Astrocytes, labelled with glial fibrilliary acidic protein (GFAP; red) and P2X7-R-IR (green) were found to co-localise in retinal flat mount. (D) Microglia labelled with mouse anti-GFP (pseudo-coloured red) from CX3CR1-GFP heterozygous mice were also found to co-localise with P2X7-R-IR (pseudo-coloured green). Scale bar, 10 microns.
    Figure Legend Snippet: (A–B) Müller cells, labelled with an antibody against glutamine synthetise (red) were found to be closely associated with P2X7-R-IR puncta in the IPL (green), but not co-localised in the IPL. (C) Astrocytes, labelled with glial fibrilliary acidic protein (GFAP; red) and P2X7-R-IR (green) were found to co-localise in retinal flat mount. (D) Microglia labelled with mouse anti-GFP (pseudo-coloured red) from CX3CR1-GFP heterozygous mice were also found to co-localise with P2X7-R-IR (pseudo-coloured green). Scale bar, 10 microns.

    Techniques Used:

    n terminal p2x7 r antibody  (Alomone Labs)


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    Alomone Labs n terminal p2x7 r antibody
    Oligonucleotide Primer List.
    N Terminal P2x7 R Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/n terminal p2x7 r antibody/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    n terminal p2x7 r antibody - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse"

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0029990

    Oligonucleotide Primer List.
    Figure Legend Snippet: Oligonucleotide Primer List.

    Techniques Used:

    Antibody List.
    Figure Legend Snippet: Antibody List.

    Techniques Used: Binding Assay, Transduction

    To assess whether P2X7-R splice variants other than the full length, variant 1 are expressed in the WT and P2X7-KO mouse retina and also whether read through of variant 1 C-terminal mRNA occurs in the P2X7-KO mouse retina, PCR for (A) the central region of the P2X7-R which is common to variants 1, 2 and 3; (B) the C-terminus from within the start of the disruption in the P2X7-KO to the end of the mRNA (variant 1 only, primer pair A); and (C) the C-terminus from after the insertion deletion in the P2X7-KO mouse to the end of the receptor (variant 1 only, primer pair B) was completed. P2X7-R variant 1 C-terminal mRNA was absent (B–C) in the P2X7-KO mouse but N-terminal mRNA variants (likely variant 2 or 3) were still expressed (A). MW, molecular weight marker and the bright bar is 300 bp. (D–E) Protein analysis of P2X7-R splice variant expression was completed using Western blots of mouse retina (D) and cortex (E). An N-terminal specific P2X7-R antibody detected the protein transcript of variant 1 mRNA in WT samples 1 and 2 but not P2X7-KO samples 3 and 4 (68.41 kDa) of both retina and cortex. Protein for variant 2/3 (50.69 kDa/49.38 kDa) and 4 (17.2 kDa) was expressed in both WT and P2X7-KO retina and cortex. A monoclonal antibody against GAPDH was applied to the same blots and indicates that similar amounts of protein were loaded for WT and P2X7-KO samples.
    Figure Legend Snippet: To assess whether P2X7-R splice variants other than the full length, variant 1 are expressed in the WT and P2X7-KO mouse retina and also whether read through of variant 1 C-terminal mRNA occurs in the P2X7-KO mouse retina, PCR for (A) the central region of the P2X7-R which is common to variants 1, 2 and 3; (B) the C-terminus from within the start of the disruption in the P2X7-KO to the end of the mRNA (variant 1 only, primer pair A); and (C) the C-terminus from after the insertion deletion in the P2X7-KO mouse to the end of the receptor (variant 1 only, primer pair B) was completed. P2X7-R variant 1 C-terminal mRNA was absent (B–C) in the P2X7-KO mouse but N-terminal mRNA variants (likely variant 2 or 3) were still expressed (A). MW, molecular weight marker and the bright bar is 300 bp. (D–E) Protein analysis of P2X7-R splice variant expression was completed using Western blots of mouse retina (D) and cortex (E). An N-terminal specific P2X7-R antibody detected the protein transcript of variant 1 mRNA in WT samples 1 and 2 but not P2X7-KO samples 3 and 4 (68.41 kDa) of both retina and cortex. Protein for variant 2/3 (50.69 kDa/49.38 kDa) and 4 (17.2 kDa) was expressed in both WT and P2X7-KO retina and cortex. A monoclonal antibody against GAPDH was applied to the same blots and indicates that similar amounts of protein were loaded for WT and P2X7-KO samples.

    Techniques Used: Variant Assay, Molecular Weight, Marker, Expressing, Western Blot

    Mice were dark adapted overnight and mixed (rod and cone) ERG responses were recorded in response to full field flashes from intensity −4.2 to 2.1 log cd.s/m 2 . (A–B) Representative mixed ERG waveforms for (A) WT and (B) P2X7-KO mice are presented. (C) There was no change in the amplitude of the rod photoreceptor derived component, a-wave, as a function of intensity. (D) The amplitude of the post-photoreceptor response, the b-wave was larger in P2X7-KO mice than in WT mice across almost all intensities tested. * indicates p<0.05, a significant difference between WT and P2X7-KO.
    Figure Legend Snippet: Mice were dark adapted overnight and mixed (rod and cone) ERG responses were recorded in response to full field flashes from intensity −4.2 to 2.1 log cd.s/m 2 . (A–B) Representative mixed ERG waveforms for (A) WT and (B) P2X7-KO mice are presented. (C) There was no change in the amplitude of the rod photoreceptor derived component, a-wave, as a function of intensity. (D) The amplitude of the post-photoreceptor response, the b-wave was larger in P2X7-KO mice than in WT mice across almost all intensities tested. * indicates p<0.05, a significant difference between WT and P2X7-KO.

    Techniques Used: Derivative Assay

    (A) Representative waveforms of the rod ERG recorded from WT (black) and P2X7-KO mice (grey) in response to a 2.1 log cd.s/m 2 intensity flash. The photoreceptor derived, rod PIII (modelled a-wave) was not altered in either (B) amplitude or (C) sensitivity in the P2X7-KO mouse when compared with the WT response. The post-photoreceptor, rod PII (modelled b-wave) was found to be (D) larger in amplitude and (E) faster to reach a maximum response. (F) The amplitude of the third of the inner retinal derived oscillatory potentials was significantly enhanced in the P2X7-KO mouse. * indicates p<0.05, a significant difference between WT and P2X7-KO.
    Figure Legend Snippet: (A) Representative waveforms of the rod ERG recorded from WT (black) and P2X7-KO mice (grey) in response to a 2.1 log cd.s/m 2 intensity flash. The photoreceptor derived, rod PIII (modelled a-wave) was not altered in either (B) amplitude or (C) sensitivity in the P2X7-KO mouse when compared with the WT response. The post-photoreceptor, rod PII (modelled b-wave) was found to be (D) larger in amplitude and (E) faster to reach a maximum response. (F) The amplitude of the third of the inner retinal derived oscillatory potentials was significantly enhanced in the P2X7-KO mouse. * indicates p<0.05, a significant difference between WT and P2X7-KO.

    Techniques Used: Derivative Assay

    (A) Representative waveforms of the cone ERG recorded from WT (black) and P2X7-KO mice (grey) in response to a 2.1 log cd.s/m 2 intensity flash. The post-photoreceptor, cone PII (modelled b-wave) was found to be larger in (B) amplitude but not faster to reach a maximum response (C) in the P2X7-KO mouse when compared with the WT response. (D) The photopic visual acuity, assessed using the optokinetic reflex to determine the spatial frequency threshold, was higher in the P2X7-KO mouse than in the WT. * indicates p<0.05, a significant difference between WT and P2X7-KO.
    Figure Legend Snippet: (A) Representative waveforms of the cone ERG recorded from WT (black) and P2X7-KO mice (grey) in response to a 2.1 log cd.s/m 2 intensity flash. The post-photoreceptor, cone PII (modelled b-wave) was found to be larger in (B) amplitude but not faster to reach a maximum response (C) in the P2X7-KO mouse when compared with the WT response. (D) The photopic visual acuity, assessed using the optokinetic reflex to determine the spatial frequency threshold, was higher in the P2X7-KO mouse than in the WT. * indicates p<0.05, a significant difference between WT and P2X7-KO.

    Techniques Used:

    Transverse sections of paraffin embedded retina from (A) WT and (B) P2X7-KO mice were stained with hematoxylin and eosin for comparison. The gross retinal morphology was similar in both mice. Scale bar, 20 microns.
    Figure Legend Snippet: Transverse sections of paraffin embedded retina from (A) WT and (B) P2X7-KO mice were stained with hematoxylin and eosin for comparison. The gross retinal morphology was similar in both mice. Scale bar, 20 microns.

    Techniques Used: Staining

    (A) Immunohistochemistry on a transverse retinal section from a WT mouse showing P2X7-R immunoreactivity in the outer plexiform layer (OPL), inner nuclear layer (INL), and ganglion cell layer (GCL). (C) The retina does not label with the P2X7-R antibody following incubation with P2X7-R specific peptide. Scale bar, 20 microns.
    Figure Legend Snippet: (A) Immunohistochemistry on a transverse retinal section from a WT mouse showing P2X7-R immunoreactivity in the outer plexiform layer (OPL), inner nuclear layer (INL), and ganglion cell layer (GCL). (C) The retina does not label with the P2X7-R antibody following incubation with P2X7-R specific peptide. Scale bar, 20 microns.

    Techniques Used: Immunohistochemistry, Incubation

    (A) P2X7-R IR (green) colocalises with horizontal cells labelled with Calbindin (red) and their terminals. (B) P2X7-R IR (green) colocalises with rod bipolar cells labelled with PKC (red). (C) Inset of rod bipolar cell terminals. (D) P2X7-R IR (green) puncta are discretely expressed within photoreceptor terminals labelled with PSD-95 (red). (E) Cone bipolar cells (Type 2 and 6) labelled with a ZNP-1 antibody (green) colocalise with P2X7-R IR (red) including both the cone bipolar cell dendrites and terminals (F). P2X7-R IR (green) colocalises with amacrine cells labelled with calretinin (G) and in particular GABAergic amacrine cells (H). (I) Ganglion cells labelled with an antibody against GFP in the Thy1-HYFP mouse (pseudo-coloured red) were P2X7-R IR (pseudo-coloured green). Scale bar, 10 microns.
    Figure Legend Snippet: (A) P2X7-R IR (green) colocalises with horizontal cells labelled with Calbindin (red) and their terminals. (B) P2X7-R IR (green) colocalises with rod bipolar cells labelled with PKC (red). (C) Inset of rod bipolar cell terminals. (D) P2X7-R IR (green) puncta are discretely expressed within photoreceptor terminals labelled with PSD-95 (red). (E) Cone bipolar cells (Type 2 and 6) labelled with a ZNP-1 antibody (green) colocalise with P2X7-R IR (red) including both the cone bipolar cell dendrites and terminals (F). P2X7-R IR (green) colocalises with amacrine cells labelled with calretinin (G) and in particular GABAergic amacrine cells (H). (I) Ganglion cells labelled with an antibody against GFP in the Thy1-HYFP mouse (pseudo-coloured red) were P2X7-R IR (pseudo-coloured green). Scale bar, 10 microns.

    Techniques Used:

    (A) P2X7-R IR (red) is associated with but does not colocalise with ribbon synapses (ribeye, green) on bipolar cell terminals (VGLUT, blue). (B) Magnified inset from A, arrows indicate ribbon synapses associated with P2X7-R IR puncta. (C) P2X7-R IR (red) colocalises with conventional synapses (bassoon, green) distinct from bipolar cell terminals (VGLUT, blue). (D) Magnified inset from C, arrows indicate conventional synapses that colocalise with P2X7-R IR puncta. Co-localisation analysis indicates that P2X7-R IR puncta do not co-localise with ribbon synapses (E) but do show significant colocalisation with conventional synapses (F) in the IPL. Scale bars are 5 microns for A & C and 2 microns for B & D.
    Figure Legend Snippet: (A) P2X7-R IR (red) is associated with but does not colocalise with ribbon synapses (ribeye, green) on bipolar cell terminals (VGLUT, blue). (B) Magnified inset from A, arrows indicate ribbon synapses associated with P2X7-R IR puncta. (C) P2X7-R IR (red) colocalises with conventional synapses (bassoon, green) distinct from bipolar cell terminals (VGLUT, blue). (D) Magnified inset from C, arrows indicate conventional synapses that colocalise with P2X7-R IR puncta. Co-localisation analysis indicates that P2X7-R IR puncta do not co-localise with ribbon synapses (E) but do show significant colocalisation with conventional synapses (F) in the IPL. Scale bars are 5 microns for A & C and 2 microns for B & D.

    Techniques Used:

    Ribbons are indicated by arrows and labelled amacine cells are indicated by astericks. (A), (B) & (C) P2X7-R immunoreactivity labels amacrine cells adjacent to rod bipolar cell ribbon synapses in the IPL. (D) A conventional amacrine cell synapse is labelled for P2X7 immunoreactivity in close proximity to a rod bipolar cell terminal. Scale bars are 200 nm for A & B and 500 nm for C & D.
    Figure Legend Snippet: Ribbons are indicated by arrows and labelled amacine cells are indicated by astericks. (A), (B) & (C) P2X7-R immunoreactivity labels amacrine cells adjacent to rod bipolar cell ribbon synapses in the IPL. (D) A conventional amacrine cell synapse is labelled for P2X7 immunoreactivity in close proximity to a rod bipolar cell terminal. Scale bars are 200 nm for A & B and 500 nm for C & D.

    Techniques Used:

    (A–B) Müller cells, labelled with an antibody against glutamine synthetise (red) were found to be closely associated with P2X7-R-IR puncta in the IPL (green), but not co-localised in the IPL. (C) Astrocytes, labelled with glial fibrilliary acidic protein (GFAP; red) and P2X7-R-IR (green) were found to co-localise in retinal flat mount. (D) Microglia labelled with mouse anti-GFP (pseudo-coloured red) from CX3CR1-GFP heterozygous mice were also found to co-localise with P2X7-R-IR (pseudo-coloured green). Scale bar, 10 microns.
    Figure Legend Snippet: (A–B) Müller cells, labelled with an antibody against glutamine synthetise (red) were found to be closely associated with P2X7-R-IR puncta in the IPL (green), but not co-localised in the IPL. (C) Astrocytes, labelled with glial fibrilliary acidic protein (GFAP; red) and P2X7-R-IR (green) were found to co-localise in retinal flat mount. (D) Microglia labelled with mouse anti-GFP (pseudo-coloured red) from CX3CR1-GFP heterozygous mice were also found to co-localise with P2X7-R-IR (pseudo-coloured green). Scale bar, 10 microns.

    Techniques Used:

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  • 95
    Alomone Labs p2x 7 apr 008
    (A) Immortalized GEC that had been depleted of <t>P2X</t> 4 or <t>P2X</t> <t>7</t> using lentiviral particles were treated with 100 µM or 3 mM ATP for 3 hours, and supernatants were collected for caspase-1 activity measurement. Caspase-1 activity was measured by ELISA as described in . (B) Total proteins isolated from GEC were subjected to immunoprecipitation (IP) by a polyclonal anti-P2X 4 antibody or Dynabeads as a control. Precipitates or total protein extract (as input) were resolved on SDS-PAGE and analyzed on immunoblots with anti-P2X 7 (top), anti-pannexin-1 (middle), or anti-P2X 4 (bottom) antibodies.
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    Alomone Labs anti purinergic p2x 7 r antibody
    (A). Astrocytes were treated with PACAP (20 ng/ml), or indirectly co-cultured with hMSCs in the absence and presence of PACAP (20 ng/ml) for 48 hours. The cultures were subjected to Q-PCR for measurement of the mRNA levels of GLT-1 and GLAST. Data are presented as mean ± SEM and expressed as the ratio of GLT-1 (GLAST) mRNA levels compared to control. * p <0.05 versus control. (B). Astrocytes were indirectly co-cultured with hMSCs in the absence or presence of PACAP for 48 hours. Membrane proteins were then extracted for western blotting using anti-GLT-1 antibody. The same blot was reprobed with <t>anti-P2X</t> 7 R antibody. <t>P2X</t> <t>7</t> <t>R</t> levels are presented as a loading control. Relative intensity of GLT-1 levels normalized to P2X 7 R was measured. Data are presented as mean ± SEM and expressed as a percentage of GLT-1 levels in the group with combinatorial treatment compared to that detected in the group co-cultured only with hMSCs. * p <0.05 versus the group only co-cultured with hMSCs. (C). Astrocytes were infected by lentivirus carrying lenti-GLT-1-GFP, and then co-cultured with hMSCs in the absence or presence of PACAP. Strong punctate fluorescence spots (arrowheads) were observed in the processes of astrocytes co-cultured with hMSCs in the presence of PACAP, when compared to that seen in astrocytes co-cultured only with hMSCs (arrows). Scale bar, 50 µm. (D). Astrocytes were treated with PACAP, or co-cultured with hMSCs in the absence or presence of PACAP. After 48 or 72 hours, the cultures were subjected to [ 3 H]-L-glutamate uptake analysis as described in . Data are presented as mean ± SEM and expressed as a ratio of the glutamate uptake in each treated group compared to that detected in the control group. * p <0.05 versus control.
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    94
    Alomone Labs affinity purified rabbit anti p2x7r antibodies
    Spleen cells from 3 to 4-mo-old MRL +/+ ( solid line ), MRL/ lpr ( dashed line ) and P2X7-deficient B6 <t>P2X7R−/−</t> ( dotted line ) mice (n = 3 mice/group) were treated for 45 min at 37°C with doses of ATP ranging from 100 to 5000 µM. Spleen cells were then triple-stained with anti-CD90, anti-CD19 and anti-CD62L mAb to assess by flow cytometry: (A) the percentage of CD62L-expressing CD19 – CD90 + T cells and (B) MFI of CD62L on CD19 – CD90 + T cells. Results are expressed as the mean percentage of initial expression ± SE.
    Affinity Purified Rabbit Anti P2x7r Antibodies, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs p2x7 receptors
    Intracellular Ca 2+ measurement (using Fura-2-AM) in macrophages subjected to treatment with the <t>P2X7</t> agonist BzATP and/or with the antagonist A740003. (a) Changes in intracellular Ca 2+ levels in macrophages from control (black line) and S. mansoni -infected (red line) mice. Inset: representative images from macrophages (control) before (baseline) and after treatment with 100 μ M BzATP. (b) Changes in intracellular Ca 2+ levels ([Ca 2+ ] i ) in macrophages from control (white bars) or S. mansoni- infected (black bars) mice. Three plates were used for each condition/animal, 10 cells/plate were chosen randomly for imaging, and cells were obtained from three animals. Data were expressed as mean and SEM. * P < 0.05; *** P < 0.001 (one-way ANOVA followed by post hoc Newman-Keuls test).
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    Alomone Labs rabbit anti p2x7r monoclonal igg
    The sequences of oligonucleotide primers.
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    Alomone Labs anti p2x7 polyclonal rabbit antibody
    Association of pannexin1 and <t>P2X7</t> in unwounded corneas and 2 hrs after injury in DiO and B6 control corneal epithelium. Proximity ligation assays were performed using antibodies to P2X7 and pannexin1. (a) Representative enface and orthogonal (ortho) images of B6 control and DiO corneas are shown. The green color displays puncta of association. Corneal epithelium was counter stained with rhodamine phalloidin (asterisk marks wound area). The numbers and letters mark the individual cells that were analyzed using CellProfiler near the wound and back from the wound. The boxes indicate the representative individual cells that are shown in (b) after PLA analysis with CellProfiler. Bar equals 20 microns (PLA enface and actin) or 15 microns (PLA ortho). (b) Analysis of overlapping puncta for each condition (unwounded, leading edge, back from leading edge) was performed and a box plot drawn using R. The mean puncta for each region are placed above the box. Mean ± SEM are plotted, and two-way ANOVA with Tukey's multiple comparison of means was performed. p < 0.05. N is a minimum of 3 independent experiments.
    Anti P2x7 Polyclonal Rabbit Antibody, 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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    Alomone Labs p2x7 r peptide
    Oligonucleotide Primer List.
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    Alomone Labs n terminal p2x7 r antibody
    Oligonucleotide Primer List.
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    (A) Immortalized GEC that had been depleted of P2X 4 or P2X 7 using lentiviral particles were treated with 100 µM or 3 mM ATP for 3 hours, and supernatants were collected for caspase-1 activity measurement. Caspase-1 activity was measured by ELISA as described in . (B) Total proteins isolated from GEC were subjected to immunoprecipitation (IP) by a polyclonal anti-P2X 4 antibody or Dynabeads as a control. Precipitates or total protein extract (as input) were resolved on SDS-PAGE and analyzed on immunoblots with anti-P2X 7 (top), anti-pannexin-1 (middle), or anti-P2X 4 (bottom) antibodies.

    Journal: PLoS ONE

    Article Title: P2X 4 Assembles with P2X 7 and Pannexin-1 in Gingival Epithelial Cells and Modulates ATP-induced Reactive Oxygen Species Production and Inflammasome Activation

    doi: 10.1371/journal.pone.0070210

    Figure Lengend Snippet: (A) Immortalized GEC that had been depleted of P2X 4 or P2X 7 using lentiviral particles were treated with 100 µM or 3 mM ATP for 3 hours, and supernatants were collected for caspase-1 activity measurement. Caspase-1 activity was measured by ELISA as described in . (B) Total proteins isolated from GEC were subjected to immunoprecipitation (IP) by a polyclonal anti-P2X 4 antibody or Dynabeads as a control. Precipitates or total protein extract (as input) were resolved on SDS-PAGE and analyzed on immunoblots with anti-P2X 7 (top), anti-pannexin-1 (middle), or anti-P2X 4 (bottom) antibodies.

    Article Snippet: Antibodies against P2X 4 (APR-002) and P2X 7 (APR-008) were obtained from Alomone Labs.

    Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Isolation, Immunoprecipitation, SDS Page, Western Blot

    Primary GEC (C and D) and immortalized GEC (A and B) were infected with or without P. gingivalis ( P.g. ) at an M.O.I. of 100 for 6 hours, followed by treatment with different pharmaceutical agents. Infected cells were treated with 100 µM ATP, 3 mM ATP, 3 mM ADP, 3 mM AMP, or 3 mM UTP individually for 1 hour (A and C). Alternatively, infected cells were pre-treated with 50 µM 5-BDBD for 15 minutes, 100 µM PPADS for 15 minutes, 100 µM oxATP for 30 minutes, or 1 mM probenecid for 10 minutes, followed by treatment with 3 mM ATP for 1 hour (B and D). The supernatants were collected and subjected to ELISA to measure IL-1β secretion. (E) Primary GEC were transfected with siRNA sequences against P2X 4 or P2X 7 for one day, and mRNA levels were detected by qPCR. (F) Primary GEC depleted of P2X 4 or P2X 7 were infected with P. gingivalis ( P.g. ) and treated with probenecid and 3 mM ATP as shown in (B). IL-1β secretion in the supernatants was analyzed by ELISA. The values showed averages and SD from duplicate samples, which were obtained from three separate experiments.

    Journal: PLoS ONE

    Article Title: P2X 4 Assembles with P2X 7 and Pannexin-1 in Gingival Epithelial Cells and Modulates ATP-induced Reactive Oxygen Species Production and Inflammasome Activation

    doi: 10.1371/journal.pone.0070210

    Figure Lengend Snippet: Primary GEC (C and D) and immortalized GEC (A and B) were infected with or without P. gingivalis ( P.g. ) at an M.O.I. of 100 for 6 hours, followed by treatment with different pharmaceutical agents. Infected cells were treated with 100 µM ATP, 3 mM ATP, 3 mM ADP, 3 mM AMP, or 3 mM UTP individually for 1 hour (A and C). Alternatively, infected cells were pre-treated with 50 µM 5-BDBD for 15 minutes, 100 µM PPADS for 15 minutes, 100 µM oxATP for 30 minutes, or 1 mM probenecid for 10 minutes, followed by treatment with 3 mM ATP for 1 hour (B and D). The supernatants were collected and subjected to ELISA to measure IL-1β secretion. (E) Primary GEC were transfected with siRNA sequences against P2X 4 or P2X 7 for one day, and mRNA levels were detected by qPCR. (F) Primary GEC depleted of P2X 4 or P2X 7 were infected with P. gingivalis ( P.g. ) and treated with probenecid and 3 mM ATP as shown in (B). IL-1β secretion in the supernatants was analyzed by ELISA. The values showed averages and SD from duplicate samples, which were obtained from three separate experiments.

    Article Snippet: Antibodies against P2X 4 (APR-002) and P2X 7 (APR-008) were obtained from Alomone Labs.

    Techniques: Infection, Enzyme-linked Immunosorbent Assay, Transfection

    Model showing the role of P2X 4 and P2X 7 in ROS production and inflammasome activation in GEC stimulated with extracellular ATP.

    Journal: PLoS ONE

    Article Title: P2X 4 Assembles with P2X 7 and Pannexin-1 in Gingival Epithelial Cells and Modulates ATP-induced Reactive Oxygen Species Production and Inflammasome Activation

    doi: 10.1371/journal.pone.0070210

    Figure Lengend Snippet: Model showing the role of P2X 4 and P2X 7 in ROS production and inflammasome activation in GEC stimulated with extracellular ATP.

    Article Snippet: Antibodies against P2X 4 (APR-002) and P2X 7 (APR-008) were obtained from Alomone Labs.

    Techniques: Activation Assay

    (A). Astrocytes were treated with PACAP (20 ng/ml), or indirectly co-cultured with hMSCs in the absence and presence of PACAP (20 ng/ml) for 48 hours. The cultures were subjected to Q-PCR for measurement of the mRNA levels of GLT-1 and GLAST. Data are presented as mean ± SEM and expressed as the ratio of GLT-1 (GLAST) mRNA levels compared to control. * p <0.05 versus control. (B). Astrocytes were indirectly co-cultured with hMSCs in the absence or presence of PACAP for 48 hours. Membrane proteins were then extracted for western blotting using anti-GLT-1 antibody. The same blot was reprobed with anti-P2X 7 R antibody. P2X 7 R levels are presented as a loading control. Relative intensity of GLT-1 levels normalized to P2X 7 R was measured. Data are presented as mean ± SEM and expressed as a percentage of GLT-1 levels in the group with combinatorial treatment compared to that detected in the group co-cultured only with hMSCs. * p <0.05 versus the group only co-cultured with hMSCs. (C). Astrocytes were infected by lentivirus carrying lenti-GLT-1-GFP, and then co-cultured with hMSCs in the absence or presence of PACAP. Strong punctate fluorescence spots (arrowheads) were observed in the processes of astrocytes co-cultured with hMSCs in the presence of PACAP, when compared to that seen in astrocytes co-cultured only with hMSCs (arrows). Scale bar, 50 µm. (D). Astrocytes were treated with PACAP, or co-cultured with hMSCs in the absence or presence of PACAP. After 48 or 72 hours, the cultures were subjected to [ 3 H]-L-glutamate uptake analysis as described in . Data are presented as mean ± SEM and expressed as a ratio of the glutamate uptake in each treated group compared to that detected in the control group. * p <0.05 versus control.

    Journal: PLoS ONE

    Article Title: Effects of Combinatorial Treatment with Pituitary Adenylate Cyclase Activating Peptide and Human Mesenchymal Stem Cells on Spinal Cord Tissue Repair

    doi: 10.1371/journal.pone.0015299

    Figure Lengend Snippet: (A). Astrocytes were treated with PACAP (20 ng/ml), or indirectly co-cultured with hMSCs in the absence and presence of PACAP (20 ng/ml) for 48 hours. The cultures were subjected to Q-PCR for measurement of the mRNA levels of GLT-1 and GLAST. Data are presented as mean ± SEM and expressed as the ratio of GLT-1 (GLAST) mRNA levels compared to control. * p <0.05 versus control. (B). Astrocytes were indirectly co-cultured with hMSCs in the absence or presence of PACAP for 48 hours. Membrane proteins were then extracted for western blotting using anti-GLT-1 antibody. The same blot was reprobed with anti-P2X 7 R antibody. P2X 7 R levels are presented as a loading control. Relative intensity of GLT-1 levels normalized to P2X 7 R was measured. Data are presented as mean ± SEM and expressed as a percentage of GLT-1 levels in the group with combinatorial treatment compared to that detected in the group co-cultured only with hMSCs. * p <0.05 versus the group only co-cultured with hMSCs. (C). Astrocytes were infected by lentivirus carrying lenti-GLT-1-GFP, and then co-cultured with hMSCs in the absence or presence of PACAP. Strong punctate fluorescence spots (arrowheads) were observed in the processes of astrocytes co-cultured with hMSCs in the presence of PACAP, when compared to that seen in astrocytes co-cultured only with hMSCs (arrows). Scale bar, 50 µm. (D). Astrocytes were treated with PACAP, or co-cultured with hMSCs in the absence or presence of PACAP. After 48 or 72 hours, the cultures were subjected to [ 3 H]-L-glutamate uptake analysis as described in . Data are presented as mean ± SEM and expressed as a ratio of the glutamate uptake in each treated group compared to that detected in the control group. * p <0.05 versus control.

    Article Snippet: The protein was identified by incubating the membrane with anti-GLT-1 antibody (1∶1000; Millipore) or anti- purinergic P2X 7 R antibody (1∶1000; Alomone Labs, Israel) overnight at 4°C, followed by HRP-conjugated secondary antibody (1∶2000) and ECL solution.

    Techniques: Cell Culture, Western Blot, Infection, Fluorescence

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

    Journal: PLoS ONE

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

    doi: 10.1371/journal.pone.0052161

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

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

    Techniques: Staining, Flow Cytometry, Expressing

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

    Journal: PLoS ONE

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

    doi: 10.1371/journal.pone.0052161

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

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

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

    Intracellular Ca 2+ measurement (using Fura-2-AM) in macrophages subjected to treatment with the P2X7 agonist BzATP and/or with the antagonist A740003. (a) Changes in intracellular Ca 2+ levels in macrophages from control (black line) and S. mansoni -infected (red line) mice. Inset: representative images from macrophages (control) before (baseline) and after treatment with 100 μ M BzATP. (b) Changes in intracellular Ca 2+ levels ([Ca 2+ ] i ) in macrophages from control (white bars) or S. mansoni- infected (black bars) mice. Three plates were used for each condition/animal, 10 cells/plate were chosen randomly for imaging, and cells were obtained from three animals. Data were expressed as mean and SEM. * P < 0.05; *** P < 0.001 (one-way ANOVA followed by post hoc Newman-Keuls test).

    Journal: Mediators of Inflammation

    Article Title: Macrophage P2X7 Receptor Function Is Reduced during Schistosomiasis: Putative Role of TGF- β 1

    doi: 10.1155/2014/134974

    Figure Lengend Snippet: Intracellular Ca 2+ measurement (using Fura-2-AM) in macrophages subjected to treatment with the P2X7 agonist BzATP and/or with the antagonist A740003. (a) Changes in intracellular Ca 2+ levels in macrophages from control (black line) and S. mansoni -infected (red line) mice. Inset: representative images from macrophages (control) before (baseline) and after treatment with 100 μ M BzATP. (b) Changes in intracellular Ca 2+ levels ([Ca 2+ ] i ) in macrophages from control (white bars) or S. mansoni- infected (black bars) mice. Three plates were used for each condition/animal, 10 cells/plate were chosen randomly for imaging, and cells were obtained from three animals. Data were expressed as mean and SEM. * P < 0.05; *** P < 0.001 (one-way ANOVA followed by post hoc Newman-Keuls test).

    Article Snippet: After three washes in PBS, samples were blocked in PBS with 10% FBS and 0.1% BSA for 30 min, washed twice in PBS, and incubated overnight with a rabbit polyclonal antibody that recognizes an extracellular epitope of P2X7 receptors (APR-008; Alomone Labs, Israel; Peptide KKGWMDPQSKGIQTGRC, corresponding to amino acid residues 136–152 of mouse P2X7 receptor; extracellular loop) diluted to 1 : 400 in PBS and 0.1% BSA.

    Techniques: Infection, Imaging

    P2X7 receptors expression in peritoneal macrophages. (a) Macrophages from uninfected (white bar) and S. mansoni -infected (black bar) mice express similar levels of P2X7 protein. n = 9 replicates from three different animals, for each group. The images above are representative Western blots (U = uninfected and I = infected). (b) P2X7 receptors expression in peritoneal macrophages from control mice (white bar, untreated) or TGF- β 1-treated cells (5 ng/mL, 24 h; black bar) are also similar. The images above are representative Western blots (U = uninfected; TGF- β 1= TGF- β 1-treated macrophages). Lysates from P2X7 KO macrophages were used as negative controls for P2X7 protein expression. Cells lysates were obtained from three animals for each group. (c) Quantitative RT-PCR (qRT-PCR) analysis of P2X7 receptor mRNA levels in peritoneal macrophages. Similar levels of P2X7 mRNA were found in uninfected (white bar), S. mansoni -infected mice (black bar), and TGF- β 1-treated (5 ng/mL, 24 h; hatched bar) mouse macrophages. N = 3–5 samples per group (using different animals).

    Journal: Mediators of Inflammation

    Article Title: Macrophage P2X7 Receptor Function Is Reduced during Schistosomiasis: Putative Role of TGF- β 1

    doi: 10.1155/2014/134974

    Figure Lengend Snippet: P2X7 receptors expression in peritoneal macrophages. (a) Macrophages from uninfected (white bar) and S. mansoni -infected (black bar) mice express similar levels of P2X7 protein. n = 9 replicates from three different animals, for each group. The images above are representative Western blots (U = uninfected and I = infected). (b) P2X7 receptors expression in peritoneal macrophages from control mice (white bar, untreated) or TGF- β 1-treated cells (5 ng/mL, 24 h; black bar) are also similar. The images above are representative Western blots (U = uninfected; TGF- β 1= TGF- β 1-treated macrophages). Lysates from P2X7 KO macrophages were used as negative controls for P2X7 protein expression. Cells lysates were obtained from three animals for each group. (c) Quantitative RT-PCR (qRT-PCR) analysis of P2X7 receptor mRNA levels in peritoneal macrophages. Similar levels of P2X7 mRNA were found in uninfected (white bar), S. mansoni -infected mice (black bar), and TGF- β 1-treated (5 ng/mL, 24 h; hatched bar) mouse macrophages. N = 3–5 samples per group (using different animals).

    Article Snippet: After three washes in PBS, samples were blocked in PBS with 10% FBS and 0.1% BSA for 30 min, washed twice in PBS, and incubated overnight with a rabbit polyclonal antibody that recognizes an extracellular epitope of P2X7 receptors (APR-008; Alomone Labs, Israel; Peptide KKGWMDPQSKGIQTGRC, corresponding to amino acid residues 136–152 of mouse P2X7 receptor; extracellular loop) diluted to 1 : 400 in PBS and 0.1% BSA.

    Techniques: Expressing, Infection, Western Blot, Quantitative RT-PCR

    Confocal microscopy analysis of P2X7 receptor expression on the cell surface of macrophages. Cells were labelled with antibodies recognizing P2X7 receptors (green) and F4/80+ (red) as well as with DAPI (blue). (a) Representative images showing macrophages from uninfected (left column), S. mansoni -infected (middle column), and TGF- β 1-treated (right column; 5 ng/mL, for 24 h) mice. The bottom row shows orthogonal slices of the representative labelled cells, with red arrows marking P2X7 expression on the plasma membrane. (b) Quantitative analysis of confocal microscopy data. The mean fluorescence intensity, showing cell surface P2X7 receptor expression in peritoneal macrophages from uninfected mice (white bar), S. mansoni -infected (black bar), and TGF- β 1-treated (hatched bar; 5 ng/mL, for 24 h) mice, was obtained from pixels intensity using ImageJ software (see methods). Inset: negative controls (macrophages incubated with secondary antibodies only). n = 6 replicates from 3 animals for each condition (*** P < 0.001 versus control, P < 0.01 TGF- β 1 versus infected group; one-way ANOVA followed by post hoc Newman-Keuls test).

    Journal: Mediators of Inflammation

    Article Title: Macrophage P2X7 Receptor Function Is Reduced during Schistosomiasis: Putative Role of TGF- β 1

    doi: 10.1155/2014/134974

    Figure Lengend Snippet: Confocal microscopy analysis of P2X7 receptor expression on the cell surface of macrophages. Cells were labelled with antibodies recognizing P2X7 receptors (green) and F4/80+ (red) as well as with DAPI (blue). (a) Representative images showing macrophages from uninfected (left column), S. mansoni -infected (middle column), and TGF- β 1-treated (right column; 5 ng/mL, for 24 h) mice. The bottom row shows orthogonal slices of the representative labelled cells, with red arrows marking P2X7 expression on the plasma membrane. (b) Quantitative analysis of confocal microscopy data. The mean fluorescence intensity, showing cell surface P2X7 receptor expression in peritoneal macrophages from uninfected mice (white bar), S. mansoni -infected (black bar), and TGF- β 1-treated (hatched bar; 5 ng/mL, for 24 h) mice, was obtained from pixels intensity using ImageJ software (see methods). Inset: negative controls (macrophages incubated with secondary antibodies only). n = 6 replicates from 3 animals for each condition (*** P < 0.001 versus control, P < 0.01 TGF- β 1 versus infected group; one-way ANOVA followed by post hoc Newman-Keuls test).

    Article Snippet: After three washes in PBS, samples were blocked in PBS with 10% FBS and 0.1% BSA for 30 min, washed twice in PBS, and incubated overnight with a rabbit polyclonal antibody that recognizes an extracellular epitope of P2X7 receptors (APR-008; Alomone Labs, Israel; Peptide KKGWMDPQSKGIQTGRC, corresponding to amino acid residues 136–152 of mouse P2X7 receptor; extracellular loop) diluted to 1 : 400 in PBS and 0.1% BSA.

    Techniques: Confocal Microscopy, Expressing, Infection, Fluorescence, Software, Incubation

    Survival curve of C57BL/6 wild type or P2X7 KO infected with approximately 80 cercariae of S. mansoni . (P2X7 KO: n = 9 females and 7 males; C57BL/6 wild type: 4 females and 10 males). *** P < 0.0001 using the Mantel-Cox log-rank test.

    Journal: Mediators of Inflammation

    Article Title: Macrophage P2X7 Receptor Function Is Reduced during Schistosomiasis: Putative Role of TGF- β 1

    doi: 10.1155/2014/134974

    Figure Lengend Snippet: Survival curve of C57BL/6 wild type or P2X7 KO infected with approximately 80 cercariae of S. mansoni . (P2X7 KO: n = 9 females and 7 males; C57BL/6 wild type: 4 females and 10 males). *** P < 0.0001 using the Mantel-Cox log-rank test.

    Article Snippet: After three washes in PBS, samples were blocked in PBS with 10% FBS and 0.1% BSA for 30 min, washed twice in PBS, and incubated overnight with a rabbit polyclonal antibody that recognizes an extracellular epitope of P2X7 receptors (APR-008; Alomone Labs, Israel; Peptide KKGWMDPQSKGIQTGRC, corresponding to amino acid residues 136–152 of mouse P2X7 receptor; extracellular loop) diluted to 1 : 400 in PBS and 0.1% BSA.

    Techniques: Infection

    The sequences of oligonucleotide primers.

    Journal: BioMed Research International

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

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: The sequences of oligonucleotide primers.

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

    Techniques:

    The antibodies for immunoblotting.

    Journal: BioMed Research International

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

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: The antibodies for immunoblotting.

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

    Techniques: Western Blot, Concentration Assay

    The antibodies for fluorescence labeling.

    Journal: BioMed Research International

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

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: The antibodies for fluorescence labeling.

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

    Techniques: Fluorescence, Labeling, Concentration Assay

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

    Journal: BioMed Research International

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

    doi: 10.1155/2020/8143754

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

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

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

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

    Journal: BioMed Research International

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

    doi: 10.1155/2020/8143754

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

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

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

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

    Journal: BioMed Research International

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

    doi: 10.1155/2020/8143754

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

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

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

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

    Journal: BioMed Research International

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

    doi: 10.1155/2020/8143754

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

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

    Techniques: Expressing, Inhibition

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

    Journal: BioMed Research International

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

    doi: 10.1155/2020/8143754

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

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

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

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

    Journal: BioMed Research International

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

    doi: 10.1155/2020/8143754

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

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

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

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

    Journal: BioMed Research International

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

    doi: 10.1155/2020/8143754

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

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

    Techniques: Inhibition

    Association of pannexin1 and P2X7 in unwounded corneas and 2 hrs after injury in DiO and B6 control corneal epithelium. Proximity ligation assays were performed using antibodies to P2X7 and pannexin1. (a) Representative enface and orthogonal (ortho) images of B6 control and DiO corneas are shown. The green color displays puncta of association. Corneal epithelium was counter stained with rhodamine phalloidin (asterisk marks wound area). The numbers and letters mark the individual cells that were analyzed using CellProfiler near the wound and back from the wound. The boxes indicate the representative individual cells that are shown in (b) after PLA analysis with CellProfiler. Bar equals 20 microns (PLA enface and actin) or 15 microns (PLA ortho). (b) Analysis of overlapping puncta for each condition (unwounded, leading edge, back from leading edge) was performed and a box plot drawn using R. The mean puncta for each region are placed above the box. Mean ± SEM are plotted, and two-way ANOVA with Tukey's multiple comparison of means was performed. p < 0.05. N is a minimum of 3 independent experiments.

    Journal: Analytical Cellular Pathology (Amsterdam)

    Article Title: Pannexin1: Role as a Sensor to Injury Is Attenuated in Pretype 2 Corneal Diabetic Epithelium

    doi: 10.1155/2021/4793338

    Figure Lengend Snippet: Association of pannexin1 and P2X7 in unwounded corneas and 2 hrs after injury in DiO and B6 control corneal epithelium. Proximity ligation assays were performed using antibodies to P2X7 and pannexin1. (a) Representative enface and orthogonal (ortho) images of B6 control and DiO corneas are shown. The green color displays puncta of association. Corneal epithelium was counter stained with rhodamine phalloidin (asterisk marks wound area). The numbers and letters mark the individual cells that were analyzed using CellProfiler near the wound and back from the wound. The boxes indicate the representative individual cells that are shown in (b) after PLA analysis with CellProfiler. Bar equals 20 microns (PLA enface and actin) or 15 microns (PLA ortho). (b) Analysis of overlapping puncta for each condition (unwounded, leading edge, back from leading edge) was performed and a box plot drawn using R. The mean puncta for each region are placed above the box. Mean ± SEM are plotted, and two-way ANOVA with Tukey's multiple comparison of means was performed. p < 0.05. N is a minimum of 3 independent experiments.

    Article Snippet: For proximity ligation assay, anti-pannexin1 monoclonal mouse antibody directed against pannexin1 (Cat. #SC-515941) was purchased from Santa Cruz Biotechnology (Dallas, TX), and anti-P2X7 polyclonal rabbit antibody targeting the extracellular domain of P2X7 receptors (Cat. #APR-008) was purchased from Alomone Labs. 10Panx, a pannexin1 mimetic inhibitory peptide (Cat. #3348), and its scrambled peptide were purchased from Tocris Biosciences (Minneapolis, MN).

    Techniques: Ligation, Staining

    Oligonucleotide Primer List.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: Oligonucleotide Primer List.

    Article Snippet: An antibody specific for the extracellular, N-terminal region of the P2X7-R peptide (Alomone labs, Cat. no#APR008) was used as it should bind all four splice variants of the P2X7-R. Retinae and cortex from both P2X7-KO and WT mice were dissected and homogenised in a HEPEs buffer containing 40 mM HEPEs, 320 mM sucrose, pH 7.5, and a protease inhibitor cocktail.

    Techniques:

    Antibody List.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: Antibody List.

    Article Snippet: An antibody specific for the extracellular, N-terminal region of the P2X7-R peptide (Alomone labs, Cat. no#APR008) was used as it should bind all four splice variants of the P2X7-R. Retinae and cortex from both P2X7-KO and WT mice were dissected and homogenised in a HEPEs buffer containing 40 mM HEPEs, 320 mM sucrose, pH 7.5, and a protease inhibitor cocktail.

    Techniques: Binding Assay, Transduction

    To assess whether P2X7-R splice variants other than the full length, variant 1 are expressed in the WT and P2X7-KO mouse retina and also whether read through of variant 1 C-terminal mRNA occurs in the P2X7-KO mouse retina, PCR for (A) the central region of the P2X7-R which is common to variants 1, 2 and 3; (B) the C-terminus from within the start of the disruption in the P2X7-KO to the end of the mRNA (variant 1 only, primer pair A); and (C) the C-terminus from after the insertion deletion in the P2X7-KO mouse to the end of the receptor (variant 1 only, primer pair B) was completed. P2X7-R variant 1 C-terminal mRNA was absent (B–C) in the P2X7-KO mouse but N-terminal mRNA variants (likely variant 2 or 3) were still expressed (A). MW, molecular weight marker and the bright bar is 300 bp. (D–E) Protein analysis of P2X7-R splice variant expression was completed using Western blots of mouse retina (D) and cortex (E). An N-terminal specific P2X7-R antibody detected the protein transcript of variant 1 mRNA in WT samples 1 and 2 but not P2X7-KO samples 3 and 4 (68.41 kDa) of both retina and cortex. Protein for variant 2/3 (50.69 kDa/49.38 kDa) and 4 (17.2 kDa) was expressed in both WT and P2X7-KO retina and cortex. A monoclonal antibody against GAPDH was applied to the same blots and indicates that similar amounts of protein were loaded for WT and P2X7-KO samples.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: To assess whether P2X7-R splice variants other than the full length, variant 1 are expressed in the WT and P2X7-KO mouse retina and also whether read through of variant 1 C-terminal mRNA occurs in the P2X7-KO mouse retina, PCR for (A) the central region of the P2X7-R which is common to variants 1, 2 and 3; (B) the C-terminus from within the start of the disruption in the P2X7-KO to the end of the mRNA (variant 1 only, primer pair A); and (C) the C-terminus from after the insertion deletion in the P2X7-KO mouse to the end of the receptor (variant 1 only, primer pair B) was completed. P2X7-R variant 1 C-terminal mRNA was absent (B–C) in the P2X7-KO mouse but N-terminal mRNA variants (likely variant 2 or 3) were still expressed (A). MW, molecular weight marker and the bright bar is 300 bp. (D–E) Protein analysis of P2X7-R splice variant expression was completed using Western blots of mouse retina (D) and cortex (E). An N-terminal specific P2X7-R antibody detected the protein transcript of variant 1 mRNA in WT samples 1 and 2 but not P2X7-KO samples 3 and 4 (68.41 kDa) of both retina and cortex. Protein for variant 2/3 (50.69 kDa/49.38 kDa) and 4 (17.2 kDa) was expressed in both WT and P2X7-KO retina and cortex. A monoclonal antibody against GAPDH was applied to the same blots and indicates that similar amounts of protein were loaded for WT and P2X7-KO samples.

    Article Snippet: An antibody specific for the extracellular, N-terminal region of the P2X7-R peptide (Alomone labs, Cat. no#APR008) was used as it should bind all four splice variants of the P2X7-R. Retinae and cortex from both P2X7-KO and WT mice were dissected and homogenised in a HEPEs buffer containing 40 mM HEPEs, 320 mM sucrose, pH 7.5, and a protease inhibitor cocktail.

    Techniques: Variant Assay, Molecular Weight, Marker, Expressing, Western Blot

    Mice were dark adapted overnight and mixed (rod and cone) ERG responses were recorded in response to full field flashes from intensity −4.2 to 2.1 log cd.s/m 2 . (A–B) Representative mixed ERG waveforms for (A) WT and (B) P2X7-KO mice are presented. (C) There was no change in the amplitude of the rod photoreceptor derived component, a-wave, as a function of intensity. (D) The amplitude of the post-photoreceptor response, the b-wave was larger in P2X7-KO mice than in WT mice across almost all intensities tested. * indicates p<0.05, a significant difference between WT and P2X7-KO.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: Mice were dark adapted overnight and mixed (rod and cone) ERG responses were recorded in response to full field flashes from intensity −4.2 to 2.1 log cd.s/m 2 . (A–B) Representative mixed ERG waveforms for (A) WT and (B) P2X7-KO mice are presented. (C) There was no change in the amplitude of the rod photoreceptor derived component, a-wave, as a function of intensity. (D) The amplitude of the post-photoreceptor response, the b-wave was larger in P2X7-KO mice than in WT mice across almost all intensities tested. * indicates p<0.05, a significant difference between WT and P2X7-KO.

    Article Snippet: An antibody specific for the extracellular, N-terminal region of the P2X7-R peptide (Alomone labs, Cat. no#APR008) was used as it should bind all four splice variants of the P2X7-R. Retinae and cortex from both P2X7-KO and WT mice were dissected and homogenised in a HEPEs buffer containing 40 mM HEPEs, 320 mM sucrose, pH 7.5, and a protease inhibitor cocktail.

    Techniques: Derivative Assay

    (A) Representative waveforms of the rod ERG recorded from WT (black) and P2X7-KO mice (grey) in response to a 2.1 log cd.s/m 2 intensity flash. The photoreceptor derived, rod PIII (modelled a-wave) was not altered in either (B) amplitude or (C) sensitivity in the P2X7-KO mouse when compared with the WT response. The post-photoreceptor, rod PII (modelled b-wave) was found to be (D) larger in amplitude and (E) faster to reach a maximum response. (F) The amplitude of the third of the inner retinal derived oscillatory potentials was significantly enhanced in the P2X7-KO mouse. * indicates p<0.05, a significant difference between WT and P2X7-KO.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: (A) Representative waveforms of the rod ERG recorded from WT (black) and P2X7-KO mice (grey) in response to a 2.1 log cd.s/m 2 intensity flash. The photoreceptor derived, rod PIII (modelled a-wave) was not altered in either (B) amplitude or (C) sensitivity in the P2X7-KO mouse when compared with the WT response. The post-photoreceptor, rod PII (modelled b-wave) was found to be (D) larger in amplitude and (E) faster to reach a maximum response. (F) The amplitude of the third of the inner retinal derived oscillatory potentials was significantly enhanced in the P2X7-KO mouse. * indicates p<0.05, a significant difference between WT and P2X7-KO.

    Article Snippet: An antibody specific for the extracellular, N-terminal region of the P2X7-R peptide (Alomone labs, Cat. no#APR008) was used as it should bind all four splice variants of the P2X7-R. Retinae and cortex from both P2X7-KO and WT mice were dissected and homogenised in a HEPEs buffer containing 40 mM HEPEs, 320 mM sucrose, pH 7.5, and a protease inhibitor cocktail.

    Techniques: Derivative Assay

    (A) Representative waveforms of the cone ERG recorded from WT (black) and P2X7-KO mice (grey) in response to a 2.1 log cd.s/m 2 intensity flash. The post-photoreceptor, cone PII (modelled b-wave) was found to be larger in (B) amplitude but not faster to reach a maximum response (C) in the P2X7-KO mouse when compared with the WT response. (D) The photopic visual acuity, assessed using the optokinetic reflex to determine the spatial frequency threshold, was higher in the P2X7-KO mouse than in the WT. * indicates p<0.05, a significant difference between WT and P2X7-KO.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: (A) Representative waveforms of the cone ERG recorded from WT (black) and P2X7-KO mice (grey) in response to a 2.1 log cd.s/m 2 intensity flash. The post-photoreceptor, cone PII (modelled b-wave) was found to be larger in (B) amplitude but not faster to reach a maximum response (C) in the P2X7-KO mouse when compared with the WT response. (D) The photopic visual acuity, assessed using the optokinetic reflex to determine the spatial frequency threshold, was higher in the P2X7-KO mouse than in the WT. * indicates p<0.05, a significant difference between WT and P2X7-KO.

    Article Snippet: An antibody specific for the extracellular, N-terminal region of the P2X7-R peptide (Alomone labs, Cat. no#APR008) was used as it should bind all four splice variants of the P2X7-R. Retinae and cortex from both P2X7-KO and WT mice were dissected and homogenised in a HEPEs buffer containing 40 mM HEPEs, 320 mM sucrose, pH 7.5, and a protease inhibitor cocktail.

    Techniques:

    Transverse sections of paraffin embedded retina from (A) WT and (B) P2X7-KO mice were stained with hematoxylin and eosin for comparison. The gross retinal morphology was similar in both mice. Scale bar, 20 microns.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: Transverse sections of paraffin embedded retina from (A) WT and (B) P2X7-KO mice were stained with hematoxylin and eosin for comparison. The gross retinal morphology was similar in both mice. Scale bar, 20 microns.

    Article Snippet: An antibody specific for the extracellular, N-terminal region of the P2X7-R peptide (Alomone labs, Cat. no#APR008) was used as it should bind all four splice variants of the P2X7-R. Retinae and cortex from both P2X7-KO and WT mice were dissected and homogenised in a HEPEs buffer containing 40 mM HEPEs, 320 mM sucrose, pH 7.5, and a protease inhibitor cocktail.

    Techniques: Staining

    (A) Immunohistochemistry on a transverse retinal section from a WT mouse showing P2X7-R immunoreactivity in the outer plexiform layer (OPL), inner nuclear layer (INL), and ganglion cell layer (GCL). (C) The retina does not label with the P2X7-R antibody following incubation with P2X7-R specific peptide. Scale bar, 20 microns.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: (A) Immunohistochemistry on a transverse retinal section from a WT mouse showing P2X7-R immunoreactivity in the outer plexiform layer (OPL), inner nuclear layer (INL), and ganglion cell layer (GCL). (C) The retina does not label with the P2X7-R antibody following incubation with P2X7-R specific peptide. Scale bar, 20 microns.

    Article Snippet: An antibody specific for the extracellular, N-terminal region of the P2X7-R peptide (Alomone labs, Cat. no#APR008) was used as it should bind all four splice variants of the P2X7-R. Retinae and cortex from both P2X7-KO and WT mice were dissected and homogenised in a HEPEs buffer containing 40 mM HEPEs, 320 mM sucrose, pH 7.5, and a protease inhibitor cocktail.

    Techniques: Immunohistochemistry, Incubation

    (A) P2X7-R IR (green) colocalises with horizontal cells labelled with Calbindin (red) and their terminals. (B) P2X7-R IR (green) colocalises with rod bipolar cells labelled with PKC (red). (C) Inset of rod bipolar cell terminals. (D) P2X7-R IR (green) puncta are discretely expressed within photoreceptor terminals labelled with PSD-95 (red). (E) Cone bipolar cells (Type 2 and 6) labelled with a ZNP-1 antibody (green) colocalise with P2X7-R IR (red) including both the cone bipolar cell dendrites and terminals (F). P2X7-R IR (green) colocalises with amacrine cells labelled with calretinin (G) and in particular GABAergic amacrine cells (H). (I) Ganglion cells labelled with an antibody against GFP in the Thy1-HYFP mouse (pseudo-coloured red) were P2X7-R IR (pseudo-coloured green). Scale bar, 10 microns.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: (A) P2X7-R IR (green) colocalises with horizontal cells labelled with Calbindin (red) and their terminals. (B) P2X7-R IR (green) colocalises with rod bipolar cells labelled with PKC (red). (C) Inset of rod bipolar cell terminals. (D) P2X7-R IR (green) puncta are discretely expressed within photoreceptor terminals labelled with PSD-95 (red). (E) Cone bipolar cells (Type 2 and 6) labelled with a ZNP-1 antibody (green) colocalise with P2X7-R IR (red) including both the cone bipolar cell dendrites and terminals (F). P2X7-R IR (green) colocalises with amacrine cells labelled with calretinin (G) and in particular GABAergic amacrine cells (H). (I) Ganglion cells labelled with an antibody against GFP in the Thy1-HYFP mouse (pseudo-coloured red) were P2X7-R IR (pseudo-coloured green). Scale bar, 10 microns.

    Article Snippet: An antibody specific for the extracellular, N-terminal region of the P2X7-R peptide (Alomone labs, Cat. no#APR008) was used as it should bind all four splice variants of the P2X7-R. Retinae and cortex from both P2X7-KO and WT mice were dissected and homogenised in a HEPEs buffer containing 40 mM HEPEs, 320 mM sucrose, pH 7.5, and a protease inhibitor cocktail.

    Techniques:

    (A) P2X7-R IR (red) is associated with but does not colocalise with ribbon synapses (ribeye, green) on bipolar cell terminals (VGLUT, blue). (B) Magnified inset from A, arrows indicate ribbon synapses associated with P2X7-R IR puncta. (C) P2X7-R IR (red) colocalises with conventional synapses (bassoon, green) distinct from bipolar cell terminals (VGLUT, blue). (D) Magnified inset from C, arrows indicate conventional synapses that colocalise with P2X7-R IR puncta. Co-localisation analysis indicates that P2X7-R IR puncta do not co-localise with ribbon synapses (E) but do show significant colocalisation with conventional synapses (F) in the IPL. Scale bars are 5 microns for A & C and 2 microns for B & D.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: (A) P2X7-R IR (red) is associated with but does not colocalise with ribbon synapses (ribeye, green) on bipolar cell terminals (VGLUT, blue). (B) Magnified inset from A, arrows indicate ribbon synapses associated with P2X7-R IR puncta. (C) P2X7-R IR (red) colocalises with conventional synapses (bassoon, green) distinct from bipolar cell terminals (VGLUT, blue). (D) Magnified inset from C, arrows indicate conventional synapses that colocalise with P2X7-R IR puncta. Co-localisation analysis indicates that P2X7-R IR puncta do not co-localise with ribbon synapses (E) but do show significant colocalisation with conventional synapses (F) in the IPL. Scale bars are 5 microns for A & C and 2 microns for B & D.

    Article Snippet: An antibody specific for the extracellular, N-terminal region of the P2X7-R peptide (Alomone labs, Cat. no#APR008) was used as it should bind all four splice variants of the P2X7-R. Retinae and cortex from both P2X7-KO and WT mice were dissected and homogenised in a HEPEs buffer containing 40 mM HEPEs, 320 mM sucrose, pH 7.5, and a protease inhibitor cocktail.

    Techniques:

    Ribbons are indicated by arrows and labelled amacine cells are indicated by astericks. (A), (B) & (C) P2X7-R immunoreactivity labels amacrine cells adjacent to rod bipolar cell ribbon synapses in the IPL. (D) A conventional amacrine cell synapse is labelled for P2X7 immunoreactivity in close proximity to a rod bipolar cell terminal. Scale bars are 200 nm for A & B and 500 nm for C & D.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: Ribbons are indicated by arrows and labelled amacine cells are indicated by astericks. (A), (B) & (C) P2X7-R immunoreactivity labels amacrine cells adjacent to rod bipolar cell ribbon synapses in the IPL. (D) A conventional amacrine cell synapse is labelled for P2X7 immunoreactivity in close proximity to a rod bipolar cell terminal. Scale bars are 200 nm for A & B and 500 nm for C & D.

    Article Snippet: An antibody specific for the extracellular, N-terminal region of the P2X7-R peptide (Alomone labs, Cat. no#APR008) was used as it should bind all four splice variants of the P2X7-R. Retinae and cortex from both P2X7-KO and WT mice were dissected and homogenised in a HEPEs buffer containing 40 mM HEPEs, 320 mM sucrose, pH 7.5, and a protease inhibitor cocktail.

    Techniques:

    (A–B) Müller cells, labelled with an antibody against glutamine synthetise (red) were found to be closely associated with P2X7-R-IR puncta in the IPL (green), but not co-localised in the IPL. (C) Astrocytes, labelled with glial fibrilliary acidic protein (GFAP; red) and P2X7-R-IR (green) were found to co-localise in retinal flat mount. (D) Microglia labelled with mouse anti-GFP (pseudo-coloured red) from CX3CR1-GFP heterozygous mice were also found to co-localise with P2X7-R-IR (pseudo-coloured green). Scale bar, 10 microns.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: (A–B) Müller cells, labelled with an antibody against glutamine synthetise (red) were found to be closely associated with P2X7-R-IR puncta in the IPL (green), but not co-localised in the IPL. (C) Astrocytes, labelled with glial fibrilliary acidic protein (GFAP; red) and P2X7-R-IR (green) were found to co-localise in retinal flat mount. (D) Microglia labelled with mouse anti-GFP (pseudo-coloured red) from CX3CR1-GFP heterozygous mice were also found to co-localise with P2X7-R-IR (pseudo-coloured green). Scale bar, 10 microns.

    Article Snippet: An antibody specific for the extracellular, N-terminal region of the P2X7-R peptide (Alomone labs, Cat. no#APR008) was used as it should bind all four splice variants of the P2X7-R. Retinae and cortex from both P2X7-KO and WT mice were dissected and homogenised in a HEPEs buffer containing 40 mM HEPEs, 320 mM sucrose, pH 7.5, and a protease inhibitor cocktail.

    Techniques:

    Oligonucleotide Primer List.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: Oligonucleotide Primer List.

    Article Snippet: The extracellular, N-terminal P2X7-R antibody directed against amino acid residues 136–152 of the receptor (Alomone labs, Cat. no#APR008) that was used for western blotting was also found to label similarly in both WT ( ) and P2X7-KO mice (data not shown).

    Techniques:

    Antibody List.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: Antibody List.

    Article Snippet: The extracellular, N-terminal P2X7-R antibody directed against amino acid residues 136–152 of the receptor (Alomone labs, Cat. no#APR008) that was used for western blotting was also found to label similarly in both WT ( ) and P2X7-KO mice (data not shown).

    Techniques: Binding Assay, Transduction

    To assess whether P2X7-R splice variants other than the full length, variant 1 are expressed in the WT and P2X7-KO mouse retina and also whether read through of variant 1 C-terminal mRNA occurs in the P2X7-KO mouse retina, PCR for (A) the central region of the P2X7-R which is common to variants 1, 2 and 3; (B) the C-terminus from within the start of the disruption in the P2X7-KO to the end of the mRNA (variant 1 only, primer pair A); and (C) the C-terminus from after the insertion deletion in the P2X7-KO mouse to the end of the receptor (variant 1 only, primer pair B) was completed. P2X7-R variant 1 C-terminal mRNA was absent (B–C) in the P2X7-KO mouse but N-terminal mRNA variants (likely variant 2 or 3) were still expressed (A). MW, molecular weight marker and the bright bar is 300 bp. (D–E) Protein analysis of P2X7-R splice variant expression was completed using Western blots of mouse retina (D) and cortex (E). An N-terminal specific P2X7-R antibody detected the protein transcript of variant 1 mRNA in WT samples 1 and 2 but not P2X7-KO samples 3 and 4 (68.41 kDa) of both retina and cortex. Protein for variant 2/3 (50.69 kDa/49.38 kDa) and 4 (17.2 kDa) was expressed in both WT and P2X7-KO retina and cortex. A monoclonal antibody against GAPDH was applied to the same blots and indicates that similar amounts of protein were loaded for WT and P2X7-KO samples.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: To assess whether P2X7-R splice variants other than the full length, variant 1 are expressed in the WT and P2X7-KO mouse retina and also whether read through of variant 1 C-terminal mRNA occurs in the P2X7-KO mouse retina, PCR for (A) the central region of the P2X7-R which is common to variants 1, 2 and 3; (B) the C-terminus from within the start of the disruption in the P2X7-KO to the end of the mRNA (variant 1 only, primer pair A); and (C) the C-terminus from after the insertion deletion in the P2X7-KO mouse to the end of the receptor (variant 1 only, primer pair B) was completed. P2X7-R variant 1 C-terminal mRNA was absent (B–C) in the P2X7-KO mouse but N-terminal mRNA variants (likely variant 2 or 3) were still expressed (A). MW, molecular weight marker and the bright bar is 300 bp. (D–E) Protein analysis of P2X7-R splice variant expression was completed using Western blots of mouse retina (D) and cortex (E). An N-terminal specific P2X7-R antibody detected the protein transcript of variant 1 mRNA in WT samples 1 and 2 but not P2X7-KO samples 3 and 4 (68.41 kDa) of both retina and cortex. Protein for variant 2/3 (50.69 kDa/49.38 kDa) and 4 (17.2 kDa) was expressed in both WT and P2X7-KO retina and cortex. A monoclonal antibody against GAPDH was applied to the same blots and indicates that similar amounts of protein were loaded for WT and P2X7-KO samples.

    Article Snippet: The extracellular, N-terminal P2X7-R antibody directed against amino acid residues 136–152 of the receptor (Alomone labs, Cat. no#APR008) that was used for western blotting was also found to label similarly in both WT ( ) and P2X7-KO mice (data not shown).

    Techniques: Variant Assay, Molecular Weight, Marker, Expressing, Western Blot

    Mice were dark adapted overnight and mixed (rod and cone) ERG responses were recorded in response to full field flashes from intensity −4.2 to 2.1 log cd.s/m 2 . (A–B) Representative mixed ERG waveforms for (A) WT and (B) P2X7-KO mice are presented. (C) There was no change in the amplitude of the rod photoreceptor derived component, a-wave, as a function of intensity. (D) The amplitude of the post-photoreceptor response, the b-wave was larger in P2X7-KO mice than in WT mice across almost all intensities tested. * indicates p<0.05, a significant difference between WT and P2X7-KO.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: Mice were dark adapted overnight and mixed (rod and cone) ERG responses were recorded in response to full field flashes from intensity −4.2 to 2.1 log cd.s/m 2 . (A–B) Representative mixed ERG waveforms for (A) WT and (B) P2X7-KO mice are presented. (C) There was no change in the amplitude of the rod photoreceptor derived component, a-wave, as a function of intensity. (D) The amplitude of the post-photoreceptor response, the b-wave was larger in P2X7-KO mice than in WT mice across almost all intensities tested. * indicates p<0.05, a significant difference between WT and P2X7-KO.

    Article Snippet: The extracellular, N-terminal P2X7-R antibody directed against amino acid residues 136–152 of the receptor (Alomone labs, Cat. no#APR008) that was used for western blotting was also found to label similarly in both WT ( ) and P2X7-KO mice (data not shown).

    Techniques: Derivative Assay

    (A) Representative waveforms of the rod ERG recorded from WT (black) and P2X7-KO mice (grey) in response to a 2.1 log cd.s/m 2 intensity flash. The photoreceptor derived, rod PIII (modelled a-wave) was not altered in either (B) amplitude or (C) sensitivity in the P2X7-KO mouse when compared with the WT response. The post-photoreceptor, rod PII (modelled b-wave) was found to be (D) larger in amplitude and (E) faster to reach a maximum response. (F) The amplitude of the third of the inner retinal derived oscillatory potentials was significantly enhanced in the P2X7-KO mouse. * indicates p<0.05, a significant difference between WT and P2X7-KO.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: (A) Representative waveforms of the rod ERG recorded from WT (black) and P2X7-KO mice (grey) in response to a 2.1 log cd.s/m 2 intensity flash. The photoreceptor derived, rod PIII (modelled a-wave) was not altered in either (B) amplitude or (C) sensitivity in the P2X7-KO mouse when compared with the WT response. The post-photoreceptor, rod PII (modelled b-wave) was found to be (D) larger in amplitude and (E) faster to reach a maximum response. (F) The amplitude of the third of the inner retinal derived oscillatory potentials was significantly enhanced in the P2X7-KO mouse. * indicates p<0.05, a significant difference between WT and P2X7-KO.

    Article Snippet: The extracellular, N-terminal P2X7-R antibody directed against amino acid residues 136–152 of the receptor (Alomone labs, Cat. no#APR008) that was used for western blotting was also found to label similarly in both WT ( ) and P2X7-KO mice (data not shown).

    Techniques: Derivative Assay

    (A) Representative waveforms of the cone ERG recorded from WT (black) and P2X7-KO mice (grey) in response to a 2.1 log cd.s/m 2 intensity flash. The post-photoreceptor, cone PII (modelled b-wave) was found to be larger in (B) amplitude but not faster to reach a maximum response (C) in the P2X7-KO mouse when compared with the WT response. (D) The photopic visual acuity, assessed using the optokinetic reflex to determine the spatial frequency threshold, was higher in the P2X7-KO mouse than in the WT. * indicates p<0.05, a significant difference between WT and P2X7-KO.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: (A) Representative waveforms of the cone ERG recorded from WT (black) and P2X7-KO mice (grey) in response to a 2.1 log cd.s/m 2 intensity flash. The post-photoreceptor, cone PII (modelled b-wave) was found to be larger in (B) amplitude but not faster to reach a maximum response (C) in the P2X7-KO mouse when compared with the WT response. (D) The photopic visual acuity, assessed using the optokinetic reflex to determine the spatial frequency threshold, was higher in the P2X7-KO mouse than in the WT. * indicates p<0.05, a significant difference between WT and P2X7-KO.

    Article Snippet: The extracellular, N-terminal P2X7-R antibody directed against amino acid residues 136–152 of the receptor (Alomone labs, Cat. no#APR008) that was used for western blotting was also found to label similarly in both WT ( ) and P2X7-KO mice (data not shown).

    Techniques:

    Transverse sections of paraffin embedded retina from (A) WT and (B) P2X7-KO mice were stained with hematoxylin and eosin for comparison. The gross retinal morphology was similar in both mice. Scale bar, 20 microns.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: Transverse sections of paraffin embedded retina from (A) WT and (B) P2X7-KO mice were stained with hematoxylin and eosin for comparison. The gross retinal morphology was similar in both mice. Scale bar, 20 microns.

    Article Snippet: The extracellular, N-terminal P2X7-R antibody directed against amino acid residues 136–152 of the receptor (Alomone labs, Cat. no#APR008) that was used for western blotting was also found to label similarly in both WT ( ) and P2X7-KO mice (data not shown).

    Techniques: Staining

    (A) Immunohistochemistry on a transverse retinal section from a WT mouse showing P2X7-R immunoreactivity in the outer plexiform layer (OPL), inner nuclear layer (INL), and ganglion cell layer (GCL). (C) The retina does not label with the P2X7-R antibody following incubation with P2X7-R specific peptide. Scale bar, 20 microns.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: (A) Immunohistochemistry on a transverse retinal section from a WT mouse showing P2X7-R immunoreactivity in the outer plexiform layer (OPL), inner nuclear layer (INL), and ganglion cell layer (GCL). (C) The retina does not label with the P2X7-R antibody following incubation with P2X7-R specific peptide. Scale bar, 20 microns.

    Article Snippet: The extracellular, N-terminal P2X7-R antibody directed against amino acid residues 136–152 of the receptor (Alomone labs, Cat. no#APR008) that was used for western blotting was also found to label similarly in both WT ( ) and P2X7-KO mice (data not shown).

    Techniques: Immunohistochemistry, Incubation

    (A) P2X7-R IR (green) colocalises with horizontal cells labelled with Calbindin (red) and their terminals. (B) P2X7-R IR (green) colocalises with rod bipolar cells labelled with PKC (red). (C) Inset of rod bipolar cell terminals. (D) P2X7-R IR (green) puncta are discretely expressed within photoreceptor terminals labelled with PSD-95 (red). (E) Cone bipolar cells (Type 2 and 6) labelled with a ZNP-1 antibody (green) colocalise with P2X7-R IR (red) including both the cone bipolar cell dendrites and terminals (F). P2X7-R IR (green) colocalises with amacrine cells labelled with calretinin (G) and in particular GABAergic amacrine cells (H). (I) Ganglion cells labelled with an antibody against GFP in the Thy1-HYFP mouse (pseudo-coloured red) were P2X7-R IR (pseudo-coloured green). Scale bar, 10 microns.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: (A) P2X7-R IR (green) colocalises with horizontal cells labelled with Calbindin (red) and their terminals. (B) P2X7-R IR (green) colocalises with rod bipolar cells labelled with PKC (red). (C) Inset of rod bipolar cell terminals. (D) P2X7-R IR (green) puncta are discretely expressed within photoreceptor terminals labelled with PSD-95 (red). (E) Cone bipolar cells (Type 2 and 6) labelled with a ZNP-1 antibody (green) colocalise with P2X7-R IR (red) including both the cone bipolar cell dendrites and terminals (F). P2X7-R IR (green) colocalises with amacrine cells labelled with calretinin (G) and in particular GABAergic amacrine cells (H). (I) Ganglion cells labelled with an antibody against GFP in the Thy1-HYFP mouse (pseudo-coloured red) were P2X7-R IR (pseudo-coloured green). Scale bar, 10 microns.

    Article Snippet: The extracellular, N-terminal P2X7-R antibody directed against amino acid residues 136–152 of the receptor (Alomone labs, Cat. no#APR008) that was used for western blotting was also found to label similarly in both WT ( ) and P2X7-KO mice (data not shown).

    Techniques:

    (A) P2X7-R IR (red) is associated with but does not colocalise with ribbon synapses (ribeye, green) on bipolar cell terminals (VGLUT, blue). (B) Magnified inset from A, arrows indicate ribbon synapses associated with P2X7-R IR puncta. (C) P2X7-R IR (red) colocalises with conventional synapses (bassoon, green) distinct from bipolar cell terminals (VGLUT, blue). (D) Magnified inset from C, arrows indicate conventional synapses that colocalise with P2X7-R IR puncta. Co-localisation analysis indicates that P2X7-R IR puncta do not co-localise with ribbon synapses (E) but do show significant colocalisation with conventional synapses (F) in the IPL. Scale bars are 5 microns for A & C and 2 microns for B & D.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: (A) P2X7-R IR (red) is associated with but does not colocalise with ribbon synapses (ribeye, green) on bipolar cell terminals (VGLUT, blue). (B) Magnified inset from A, arrows indicate ribbon synapses associated with P2X7-R IR puncta. (C) P2X7-R IR (red) colocalises with conventional synapses (bassoon, green) distinct from bipolar cell terminals (VGLUT, blue). (D) Magnified inset from C, arrows indicate conventional synapses that colocalise with P2X7-R IR puncta. Co-localisation analysis indicates that P2X7-R IR puncta do not co-localise with ribbon synapses (E) but do show significant colocalisation with conventional synapses (F) in the IPL. Scale bars are 5 microns for A & C and 2 microns for B & D.

    Article Snippet: The extracellular, N-terminal P2X7-R antibody directed against amino acid residues 136–152 of the receptor (Alomone labs, Cat. no#APR008) that was used for western blotting was also found to label similarly in both WT ( ) and P2X7-KO mice (data not shown).

    Techniques:

    Ribbons are indicated by arrows and labelled amacine cells are indicated by astericks. (A), (B) & (C) P2X7-R immunoreactivity labels amacrine cells adjacent to rod bipolar cell ribbon synapses in the IPL. (D) A conventional amacrine cell synapse is labelled for P2X7 immunoreactivity in close proximity to a rod bipolar cell terminal. Scale bars are 200 nm for A & B and 500 nm for C & D.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: Ribbons are indicated by arrows and labelled amacine cells are indicated by astericks. (A), (B) & (C) P2X7-R immunoreactivity labels amacrine cells adjacent to rod bipolar cell ribbon synapses in the IPL. (D) A conventional amacrine cell synapse is labelled for P2X7 immunoreactivity in close proximity to a rod bipolar cell terminal. Scale bars are 200 nm for A & B and 500 nm for C & D.

    Article Snippet: The extracellular, N-terminal P2X7-R antibody directed against amino acid residues 136–152 of the receptor (Alomone labs, Cat. no#APR008) that was used for western blotting was also found to label similarly in both WT ( ) and P2X7-KO mice (data not shown).

    Techniques:

    (A–B) Müller cells, labelled with an antibody against glutamine synthetise (red) were found to be closely associated with P2X7-R-IR puncta in the IPL (green), but not co-localised in the IPL. (C) Astrocytes, labelled with glial fibrilliary acidic protein (GFAP; red) and P2X7-R-IR (green) were found to co-localise in retinal flat mount. (D) Microglia labelled with mouse anti-GFP (pseudo-coloured red) from CX3CR1-GFP heterozygous mice were also found to co-localise with P2X7-R-IR (pseudo-coloured green). Scale bar, 10 microns.

    Journal: PLoS ONE

    Article Title: Rod and Cone Pathway Signalling Is Altered in the P2X7 Receptor Knock Out Mouse

    doi: 10.1371/journal.pone.0029990

    Figure Lengend Snippet: (A–B) Müller cells, labelled with an antibody against glutamine synthetise (red) were found to be closely associated with P2X7-R-IR puncta in the IPL (green), but not co-localised in the IPL. (C) Astrocytes, labelled with glial fibrilliary acidic protein (GFAP; red) and P2X7-R-IR (green) were found to co-localise in retinal flat mount. (D) Microglia labelled with mouse anti-GFP (pseudo-coloured red) from CX3CR1-GFP heterozygous mice were also found to co-localise with P2X7-R-IR (pseudo-coloured green). Scale bar, 10 microns.

    Article Snippet: The extracellular, N-terminal P2X7-R antibody directed against amino acid residues 136–152 of the receptor (Alomone labs, Cat. no#APR008) that was used for western blotting was also found to label similarly in both WT ( ) and P2X7-KO mice (data not shown).

    Techniques: