p2x 7 apr 008  (Alomone Labs)


Bioz Verified Symbol Alomone Labs is a verified supplier
Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95

    Structured Review

    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, 2023-01
    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

    rabbit anti p2x7  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Alomone Labs rabbit anti p2x7
    Effect of Schisandrin B on expression levels of <t>P2X7</t> receptor in SCG of DM rats. (a) The schedule of experiment process. (b) The protein level of P2X7 protein in all group was measured by Western blotting. (c) The relative expression level of P2X7 protein to individual β -actin internal control was shown. Data are mean ± SD from four independent experiments. ( ∗ P < 0.05, ∗∗ P < 0.01 compared with the control group; # P < 0.05 compared with the DM group).
    Rabbit Anti P2x7, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti p2x7/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti p2x7 - by Bioz Stars, 2023-01
    86/100 stars

    Images

    1) Product Images from "Schisandrin B Alleviates Diabetic Cardiac Autonomic neuropathy Induced by P2X7 Receptor in Superior Cervical Ganglion via NLRP3"

    Article Title: Schisandrin B Alleviates Diabetic Cardiac Autonomic neuropathy Induced by P2X7 Receptor in Superior Cervical Ganglion via NLRP3

    Journal: Disease Markers

    doi: 10.1155/2023/9956950

    Effect of Schisandrin B on expression levels of P2X7 receptor in SCG of DM rats. (a) The schedule of experiment process. (b) The protein level of P2X7 protein in all group was measured by Western blotting. (c) The relative expression level of P2X7 protein to individual β -actin internal control was shown. Data are mean ± SD from four independent experiments. ( ∗ P < 0.05, ∗∗ P < 0.01 compared with the control group; # P < 0.05 compared with the DM group).
    Figure Legend Snippet: Effect of Schisandrin B on expression levels of P2X7 receptor in SCG of DM rats. (a) The schedule of experiment process. (b) The protein level of P2X7 protein in all group was measured by Western blotting. (c) The relative expression level of P2X7 protein to individual β -actin internal control was shown. Data are mean ± SD from four independent experiments. ( ∗ P < 0.05, ∗∗ P < 0.01 compared with the control group; # P < 0.05 compared with the DM group).

    Techniques Used: Expressing, Western Blot

    Effect of Schisandrin B on heart rate and blood pressure in rats.
    Figure Legend Snippet: Effect of Schisandrin B on heart rate and blood pressure in rats.

    Techniques Used: shRNA

    Effects of Schisandrin B on heart rate variability in rats.
    Figure Legend Snippet: Effects of Schisandrin B on heart rate variability in rats.

    Techniques Used: shRNA

    The double-label immunofluorescence staining of P2X7 and GS in SCG. The expression of P2X7 receptor and GS in SCG was detected by double-label immunofluorescence staining. The green signal represents GS staining with FITC, and the red signal indicates P2X7 staining with TRITC. The merged image represents the double staining of P2X7 and GS. Scale bar, 20 μ m.
    Figure Legend Snippet: The double-label immunofluorescence staining of P2X7 and GS in SCG. The expression of P2X7 receptor and GS in SCG was detected by double-label immunofluorescence staining. The green signal represents GS staining with FITC, and the red signal indicates P2X7 staining with TRITC. The merged image represents the double staining of P2X7 and GS. Scale bar, 20 μ m.

    Techniques Used: Immunofluorescence, Staining, Expressing, Double Staining

    p2x7 c term  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Alomone Labs p2x7 c term
    <t>P2X7</t> −/− exerted protection against NaIO 3 -induced photoreceptor injury. ( A ) Representative scotopic ERG responses from WT and P2X7 −/− mice with or without NaIO 3 (25 mg/kg) intraperitoneal injection. Three days after injection, ERG analysis was performed. ( B ) Quantification of the average amplitudes of a and b waves from WT control mice ( n = 8), WT NaIO 3 -treated mice ( n = 8), P2X7 −/− control mice ( n = 8) and P2X7 −/− NaIO 3 -treated mice ( n = 10). ( C ) Quantification of the implicit time of a and b waves from the 4 groups of mice. The data were presented as the mean ± SEM. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .
    P2x7 C Term, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p2x7 c term/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p2x7 c term - by Bioz Stars, 2023-01
    86/100 stars

    Images

    1) Product Images from "P2X7 Is Involved in the Mouse Retinal Degeneration via the Coordinated Actions in Different Retinal Cell Types"

    Article Title: P2X7 Is Involved in the Mouse Retinal Degeneration via the Coordinated Actions in Different Retinal Cell Types

    Journal: Antioxidants

    doi: 10.3390/antiox12010141

    P2X7 −/− exerted protection against NaIO 3 -induced photoreceptor injury. ( A ) Representative scotopic ERG responses from WT and P2X7 −/− mice with or without NaIO 3 (25 mg/kg) intraperitoneal injection. Three days after injection, ERG analysis was performed. ( B ) Quantification of the average amplitudes of a and b waves from WT control mice ( n = 8), WT NaIO 3 -treated mice ( n = 8), P2X7 −/− control mice ( n = 8) and P2X7 −/− NaIO 3 -treated mice ( n = 10). ( C ) Quantification of the implicit time of a and b waves from the 4 groups of mice. The data were presented as the mean ± SEM. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .
    Figure Legend Snippet: P2X7 −/− exerted protection against NaIO 3 -induced photoreceptor injury. ( A ) Representative scotopic ERG responses from WT and P2X7 −/− mice with or without NaIO 3 (25 mg/kg) intraperitoneal injection. Three days after injection, ERG analysis was performed. ( B ) Quantification of the average amplitudes of a and b waves from WT control mice ( n = 8), WT NaIO 3 -treated mice ( n = 8), P2X7 −/− control mice ( n = 8) and P2X7 −/− NaIO 3 -treated mice ( n = 10). ( C ) Quantification of the implicit time of a and b waves from the 4 groups of mice. The data were presented as the mean ± SEM. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Techniques Used: Injection

    P2X7 knockout protected mice from NaIO 3 -induced retinal epithelium injury. OCT images in posterior pole region ( A ) and optical nerve area ( B ) revealed retinal pigment epithelium degeneration after the mice were treated with 25 mg/kg of NaIO 3 for 3 days. RPE damage (yellow arrow in ( A , B ) was observed in the WT group but not in the P2X7 −/− group treated with NaIO 3 . Hyperreflective spots in both the vitreous region (blue arrow in ( B )) and the retina (orange arrow in ( B ) were shown. The ONL thickness was marked by the red double arrowheads in ( B ). The mice numbers were 8, 8, 8 and 10 for WT control, WT NaIO 3 , P2X7 −/− control and P2X7 −/− NaIO 3 groups, respectively. Scale bar: 200 μm. RPE, retinal pigment epithelium; OS, outer segment; IS, inner segment; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer.
    Figure Legend Snippet: P2X7 knockout protected mice from NaIO 3 -induced retinal epithelium injury. OCT images in posterior pole region ( A ) and optical nerve area ( B ) revealed retinal pigment epithelium degeneration after the mice were treated with 25 mg/kg of NaIO 3 for 3 days. RPE damage (yellow arrow in ( A , B ) was observed in the WT group but not in the P2X7 −/− group treated with NaIO 3 . Hyperreflective spots in both the vitreous region (blue arrow in ( B )) and the retina (orange arrow in ( B ) were shown. The ONL thickness was marked by the red double arrowheads in ( B ). The mice numbers were 8, 8, 8 and 10 for WT control, WT NaIO 3 , P2X7 −/− control and P2X7 −/− NaIO 3 groups, respectively. Scale bar: 200 μm. RPE, retinal pigment epithelium; OS, outer segment; IS, inner segment; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer.

    Techniques Used: Knock-Out

    Histological H&E examination of retinal injury caused by NaIO 3 was alleviated in P2X7 −/− mice. Retinal morphology and loss of photoreceptors and cell nuclei were evaluated by H&E by light microscopy at 3 days post-intraperitoneal injection of NaIO 3 (25 mg/kg) in WT and P2X7 −/− mice. Shown were representative light microscopic images of H&E staining in retinal sections. Enlargement of cell bodies in the ONL and INL, pigment granules along the RPE layer (black arrows) and ONL thinning were observed. Quantitative analysis of ONL thickness was shown. The mice number was 12 for each group. Scale bar: 200 μm. * p < 0.05, indicating significant decrease in ONL thickness by NaIO 3. # p < 0.05 compared with the WT control group treated with NaIO 3 .
    Figure Legend Snippet: Histological H&E examination of retinal injury caused by NaIO 3 was alleviated in P2X7 −/− mice. Retinal morphology and loss of photoreceptors and cell nuclei were evaluated by H&E by light microscopy at 3 days post-intraperitoneal injection of NaIO 3 (25 mg/kg) in WT and P2X7 −/− mice. Shown were representative light microscopic images of H&E staining in retinal sections. Enlargement of cell bodies in the ONL and INL, pigment granules along the RPE layer (black arrows) and ONL thinning were observed. Quantitative analysis of ONL thickness was shown. The mice number was 12 for each group. Scale bar: 200 μm. * p < 0.05, indicating significant decrease in ONL thickness by NaIO 3. # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Techniques Used: Light Microscopy, Injection, Staining

    Histological examination of PAS and Masson trichrome staining. Representative light microscope images of PAS ( A ) and Masson trichrome ( B ) staining of retinal sections in WT and P2X7 −/− mice at 3 days post-intraperitoneal injection of NaIO 3 (25 mg/kg) were shown. The visible retinal pigment granules were shown as black arrows in both PAS ( A ) and Masson trichrome ( B ) staining. The mice number for PAS staining was 5 for each group and for Masson trichrome staining was 7 for each group. Scale bar: 200 μm.
    Figure Legend Snippet: Histological examination of PAS and Masson trichrome staining. Representative light microscope images of PAS ( A ) and Masson trichrome ( B ) staining of retinal sections in WT and P2X7 −/− mice at 3 days post-intraperitoneal injection of NaIO 3 (25 mg/kg) were shown. The visible retinal pigment granules were shown as black arrows in both PAS ( A ) and Masson trichrome ( B ) staining. The mice number for PAS staining was 5 for each group and for Masson trichrome staining was 7 for each group. Scale bar: 200 μm.

    Techniques Used: Staining, Light Microscopy, Injection

    NaIO 3 -induced neuroinflammation in retina was reversed by P2X7 knockout. After 36 h post-injection of NaIO 3 (25 mg/kg), mice retina was collected. ( A ) Total mRNA was extracted and reversely transcribed for quantitative PCR analysis of NLRP3, IL-1β and IL-6 mRNA expression. The mice number for qPCR analysis was 8 for each group. Values were normalized to β-actin gene expression and were expressed relative to the control group. ( B ) Parallelly, serum from mice was collected and subjected to ELISA to determine the amounts of IL-1β, TNF-α and IL-6, respectively. The mice number for ELISA was 5 for each group. The data were presented as the mean ± SEM. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .
    Figure Legend Snippet: NaIO 3 -induced neuroinflammation in retina was reversed by P2X7 knockout. After 36 h post-injection of NaIO 3 (25 mg/kg), mice retina was collected. ( A ) Total mRNA was extracted and reversely transcribed for quantitative PCR analysis of NLRP3, IL-1β and IL-6 mRNA expression. The mice number for qPCR analysis was 8 for each group. Values were normalized to β-actin gene expression and were expressed relative to the control group. ( B ) Parallelly, serum from mice was collected and subjected to ELISA to determine the amounts of IL-1β, TNF-α and IL-6, respectively. The mice number for ELISA was 5 for each group. The data were presented as the mean ± SEM. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Techniques Used: Knock-Out, Injection, Real-time Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay

    NaIO 3 -induced caspase expression in retina was reversed by P2X7 knockout . ( A ) After 3 days post-injection of NaIO 3 (25 mg/kg), mice retina was collected, and total mRNA was extracted and reversely transcribed for quantitative PCR analyses of caspase-3, caspase-7 and caspase-8 mRNA expression. The mice number for qPCR analysis was 5 for each group. Values were normalized to β-actin gene expression and were expressed relative to the control group. ( B ) Using the retinal tissues to measure the cellular contents of total NADP, NADPH and NADP/NADPH as per manufacturer’s instructions. The mice number for NADPH activity was 5 for each group. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .
    Figure Legend Snippet: NaIO 3 -induced caspase expression in retina was reversed by P2X7 knockout . ( A ) After 3 days post-injection of NaIO 3 (25 mg/kg), mice retina was collected, and total mRNA was extracted and reversely transcribed for quantitative PCR analyses of caspase-3, caspase-7 and caspase-8 mRNA expression. The mice number for qPCR analysis was 5 for each group. Values were normalized to β-actin gene expression and were expressed relative to the control group. ( B ) Using the retinal tissues to measure the cellular contents of total NADP, NADPH and NADP/NADPH as per manufacturer’s instructions. The mice number for NADPH activity was 5 for each group. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Techniques Used: Expressing, Knock-Out, Injection, Real-time Polymerase Chain Reaction, Activity Assay

    Comparison of P2X7 expression and effects of BzATP on cell viability in BV-2, 661W, ARPE-19 and rMC1 cells. ( A ) P2X7 protein level in each cell type was determined by immunoblotting. ( B , C ) Cells were treated with NaIO 3 (30 mM) for the indicated times. P2X7 mRNA level was determined by real-time PCR ( B ) and P2X7 protein was determined by immunoblotting ( C ). ( D ) Each cell type was treated with vehicle or BzATP (200 μM) for 6 or 18 h. Cell death was determined by Annexin V/PI staining with FACS. * p < 0.05, indicating the increased P2X7 gene expression by NaIO 3 ( B ) and the cell death caused by BzATP ( D ).
    Figure Legend Snippet: Comparison of P2X7 expression and effects of BzATP on cell viability in BV-2, 661W, ARPE-19 and rMC1 cells. ( A ) P2X7 protein level in each cell type was determined by immunoblotting. ( B , C ) Cells were treated with NaIO 3 (30 mM) for the indicated times. P2X7 mRNA level was determined by real-time PCR ( B ) and P2X7 protein was determined by immunoblotting ( C ). ( D ) Each cell type was treated with vehicle or BzATP (200 μM) for 6 or 18 h. Cell death was determined by Annexin V/PI staining with FACS. * p < 0.05, indicating the increased P2X7 gene expression by NaIO 3 ( B ) and the cell death caused by BzATP ( D ).

    Techniques Used: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Staining

    Schematic representation of P2X7 activation in amplification of NaIO 3 -induced neuroinflammation and retinal cell death. ( Left ) In normal conditions, P2X7 is constitutively and highly expressed in microglia than photoreceptors, RPE and Müller cells. ( Right ) Upon NaIO 3 treatment, P2X7 expression in photoreceptors, Müller cells and RPE cells is increased, and cell death in these cell types, as well as microglia, is evoked. ATP releasing from the dying and damaged cells acts as a DAMP to activate microglia for inflammatory IL-1β and IL-6 gene expression. High concentration of ATP might also promote RPE cell death under NaIO 3 stress.
    Figure Legend Snippet: Schematic representation of P2X7 activation in amplification of NaIO 3 -induced neuroinflammation and retinal cell death. ( Left ) In normal conditions, P2X7 is constitutively and highly expressed in microglia than photoreceptors, RPE and Müller cells. ( Right ) Upon NaIO 3 treatment, P2X7 expression in photoreceptors, Müller cells and RPE cells is increased, and cell death in these cell types, as well as microglia, is evoked. ATP releasing from the dying and damaged cells acts as a DAMP to activate microglia for inflammatory IL-1β and IL-6 gene expression. High concentration of ATP might also promote RPE cell death under NaIO 3 stress.

    Techniques Used: Activation Assay, Amplification, Expressing, Concentration Assay

    p2x7 extracellular ecto  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Alomone Labs p2x7 extracellular ecto
    <t>P2X7</t> −/− exerted protection against NaIO 3 -induced photoreceptor injury. ( A ) Representative scotopic ERG responses from WT and P2X7 −/− mice with or without NaIO 3 (25 mg/kg) intraperitoneal injection. Three days after injection, ERG analysis was performed. ( B ) Quantification of the average amplitudes of a and b waves from WT control mice ( n = 8), WT NaIO 3 -treated mice ( n = 8), P2X7 −/− control mice ( n = 8) and P2X7 −/− NaIO 3 -treated mice ( n = 10). ( C ) Quantification of the implicit time of a and b waves from the 4 groups of mice. The data were presented as the mean ± SEM. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .
    P2x7 Extracellular Ecto, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p2x7 extracellular ecto/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p2x7 extracellular ecto - by Bioz Stars, 2023-01
    86/100 stars

    Images

    1) Product Images from "P2X7 Is Involved in the Mouse Retinal Degeneration via the Coordinated Actions in Different Retinal Cell Types"

    Article Title: P2X7 Is Involved in the Mouse Retinal Degeneration via the Coordinated Actions in Different Retinal Cell Types

    Journal: Antioxidants

    doi: 10.3390/antiox12010141

    P2X7 −/− exerted protection against NaIO 3 -induced photoreceptor injury. ( A ) Representative scotopic ERG responses from WT and P2X7 −/− mice with or without NaIO 3 (25 mg/kg) intraperitoneal injection. Three days after injection, ERG analysis was performed. ( B ) Quantification of the average amplitudes of a and b waves from WT control mice ( n = 8), WT NaIO 3 -treated mice ( n = 8), P2X7 −/− control mice ( n = 8) and P2X7 −/− NaIO 3 -treated mice ( n = 10). ( C ) Quantification of the implicit time of a and b waves from the 4 groups of mice. The data were presented as the mean ± SEM. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .
    Figure Legend Snippet: P2X7 −/− exerted protection against NaIO 3 -induced photoreceptor injury. ( A ) Representative scotopic ERG responses from WT and P2X7 −/− mice with or without NaIO 3 (25 mg/kg) intraperitoneal injection. Three days after injection, ERG analysis was performed. ( B ) Quantification of the average amplitudes of a and b waves from WT control mice ( n = 8), WT NaIO 3 -treated mice ( n = 8), P2X7 −/− control mice ( n = 8) and P2X7 −/− NaIO 3 -treated mice ( n = 10). ( C ) Quantification of the implicit time of a and b waves from the 4 groups of mice. The data were presented as the mean ± SEM. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Techniques Used: Injection

    P2X7 knockout protected mice from NaIO 3 -induced retinal epithelium injury. OCT images in posterior pole region ( A ) and optical nerve area ( B ) revealed retinal pigment epithelium degeneration after the mice were treated with 25 mg/kg of NaIO 3 for 3 days. RPE damage (yellow arrow in ( A , B ) was observed in the WT group but not in the P2X7 −/− group treated with NaIO 3 . Hyperreflective spots in both the vitreous region (blue arrow in ( B )) and the retina (orange arrow in ( B ) were shown. The ONL thickness was marked by the red double arrowheads in ( B ). The mice numbers were 8, 8, 8 and 10 for WT control, WT NaIO 3 , P2X7 −/− control and P2X7 −/− NaIO 3 groups, respectively. Scale bar: 200 μm. RPE, retinal pigment epithelium; OS, outer segment; IS, inner segment; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer.
    Figure Legend Snippet: P2X7 knockout protected mice from NaIO 3 -induced retinal epithelium injury. OCT images in posterior pole region ( A ) and optical nerve area ( B ) revealed retinal pigment epithelium degeneration after the mice were treated with 25 mg/kg of NaIO 3 for 3 days. RPE damage (yellow arrow in ( A , B ) was observed in the WT group but not in the P2X7 −/− group treated with NaIO 3 . Hyperreflective spots in both the vitreous region (blue arrow in ( B )) and the retina (orange arrow in ( B ) were shown. The ONL thickness was marked by the red double arrowheads in ( B ). The mice numbers were 8, 8, 8 and 10 for WT control, WT NaIO 3 , P2X7 −/− control and P2X7 −/− NaIO 3 groups, respectively. Scale bar: 200 μm. RPE, retinal pigment epithelium; OS, outer segment; IS, inner segment; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer.

    Techniques Used: Knock-Out

    Histological H&E examination of retinal injury caused by NaIO 3 was alleviated in P2X7 −/− mice. Retinal morphology and loss of photoreceptors and cell nuclei were evaluated by H&E by light microscopy at 3 days post-intraperitoneal injection of NaIO 3 (25 mg/kg) in WT and P2X7 −/− mice. Shown were representative light microscopic images of H&E staining in retinal sections. Enlargement of cell bodies in the ONL and INL, pigment granules along the RPE layer (black arrows) and ONL thinning were observed. Quantitative analysis of ONL thickness was shown. The mice number was 12 for each group. Scale bar: 200 μm. * p < 0.05, indicating significant decrease in ONL thickness by NaIO 3. # p < 0.05 compared with the WT control group treated with NaIO 3 .
    Figure Legend Snippet: Histological H&E examination of retinal injury caused by NaIO 3 was alleviated in P2X7 −/− mice. Retinal morphology and loss of photoreceptors and cell nuclei were evaluated by H&E by light microscopy at 3 days post-intraperitoneal injection of NaIO 3 (25 mg/kg) in WT and P2X7 −/− mice. Shown were representative light microscopic images of H&E staining in retinal sections. Enlargement of cell bodies in the ONL and INL, pigment granules along the RPE layer (black arrows) and ONL thinning were observed. Quantitative analysis of ONL thickness was shown. The mice number was 12 for each group. Scale bar: 200 μm. * p < 0.05, indicating significant decrease in ONL thickness by NaIO 3. # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Techniques Used: Light Microscopy, Injection, Staining

    Histological examination of PAS and Masson trichrome staining. Representative light microscope images of PAS ( A ) and Masson trichrome ( B ) staining of retinal sections in WT and P2X7 −/− mice at 3 days post-intraperitoneal injection of NaIO 3 (25 mg/kg) were shown. The visible retinal pigment granules were shown as black arrows in both PAS ( A ) and Masson trichrome ( B ) staining. The mice number for PAS staining was 5 for each group and for Masson trichrome staining was 7 for each group. Scale bar: 200 μm.
    Figure Legend Snippet: Histological examination of PAS and Masson trichrome staining. Representative light microscope images of PAS ( A ) and Masson trichrome ( B ) staining of retinal sections in WT and P2X7 −/− mice at 3 days post-intraperitoneal injection of NaIO 3 (25 mg/kg) were shown. The visible retinal pigment granules were shown as black arrows in both PAS ( A ) and Masson trichrome ( B ) staining. The mice number for PAS staining was 5 for each group and for Masson trichrome staining was 7 for each group. Scale bar: 200 μm.

    Techniques Used: Staining, Light Microscopy, Injection

    NaIO 3 -induced neuroinflammation in retina was reversed by P2X7 knockout. After 36 h post-injection of NaIO 3 (25 mg/kg), mice retina was collected. ( A ) Total mRNA was extracted and reversely transcribed for quantitative PCR analysis of NLRP3, IL-1β and IL-6 mRNA expression. The mice number for qPCR analysis was 8 for each group. Values were normalized to β-actin gene expression and were expressed relative to the control group. ( B ) Parallelly, serum from mice was collected and subjected to ELISA to determine the amounts of IL-1β, TNF-α and IL-6, respectively. The mice number for ELISA was 5 for each group. The data were presented as the mean ± SEM. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .
    Figure Legend Snippet: NaIO 3 -induced neuroinflammation in retina was reversed by P2X7 knockout. After 36 h post-injection of NaIO 3 (25 mg/kg), mice retina was collected. ( A ) Total mRNA was extracted and reversely transcribed for quantitative PCR analysis of NLRP3, IL-1β and IL-6 mRNA expression. The mice number for qPCR analysis was 8 for each group. Values were normalized to β-actin gene expression and were expressed relative to the control group. ( B ) Parallelly, serum from mice was collected and subjected to ELISA to determine the amounts of IL-1β, TNF-α and IL-6, respectively. The mice number for ELISA was 5 for each group. The data were presented as the mean ± SEM. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Techniques Used: Knock-Out, Injection, Real-time Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay

    NaIO 3 -induced caspase expression in retina was reversed by P2X7 knockout . ( A ) After 3 days post-injection of NaIO 3 (25 mg/kg), mice retina was collected, and total mRNA was extracted and reversely transcribed for quantitative PCR analyses of caspase-3, caspase-7 and caspase-8 mRNA expression. The mice number for qPCR analysis was 5 for each group. Values were normalized to β-actin gene expression and were expressed relative to the control group. ( B ) Using the retinal tissues to measure the cellular contents of total NADP, NADPH and NADP/NADPH as per manufacturer’s instructions. The mice number for NADPH activity was 5 for each group. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .
    Figure Legend Snippet: NaIO 3 -induced caspase expression in retina was reversed by P2X7 knockout . ( A ) After 3 days post-injection of NaIO 3 (25 mg/kg), mice retina was collected, and total mRNA was extracted and reversely transcribed for quantitative PCR analyses of caspase-3, caspase-7 and caspase-8 mRNA expression. The mice number for qPCR analysis was 5 for each group. Values were normalized to β-actin gene expression and were expressed relative to the control group. ( B ) Using the retinal tissues to measure the cellular contents of total NADP, NADPH and NADP/NADPH as per manufacturer’s instructions. The mice number for NADPH activity was 5 for each group. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Techniques Used: Expressing, Knock-Out, Injection, Real-time Polymerase Chain Reaction, Activity Assay

    Comparison of P2X7 expression and effects of BzATP on cell viability in BV-2, 661W, ARPE-19 and rMC1 cells. ( A ) P2X7 protein level in each cell type was determined by immunoblotting. ( B , C ) Cells were treated with NaIO 3 (30 mM) for the indicated times. P2X7 mRNA level was determined by real-time PCR ( B ) and P2X7 protein was determined by immunoblotting ( C ). ( D ) Each cell type was treated with vehicle or BzATP (200 μM) for 6 or 18 h. Cell death was determined by Annexin V/PI staining with FACS. * p < 0.05, indicating the increased P2X7 gene expression by NaIO 3 ( B ) and the cell death caused by BzATP ( D ).
    Figure Legend Snippet: Comparison of P2X7 expression and effects of BzATP on cell viability in BV-2, 661W, ARPE-19 and rMC1 cells. ( A ) P2X7 protein level in each cell type was determined by immunoblotting. ( B , C ) Cells were treated with NaIO 3 (30 mM) for the indicated times. P2X7 mRNA level was determined by real-time PCR ( B ) and P2X7 protein was determined by immunoblotting ( C ). ( D ) Each cell type was treated with vehicle or BzATP (200 μM) for 6 or 18 h. Cell death was determined by Annexin V/PI staining with FACS. * p < 0.05, indicating the increased P2X7 gene expression by NaIO 3 ( B ) and the cell death caused by BzATP ( D ).

    Techniques Used: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Staining

    Schematic representation of P2X7 activation in amplification of NaIO 3 -induced neuroinflammation and retinal cell death. ( Left ) In normal conditions, P2X7 is constitutively and highly expressed in microglia than photoreceptors, RPE and Müller cells. ( Right ) Upon NaIO 3 treatment, P2X7 expression in photoreceptors, Müller cells and RPE cells is increased, and cell death in these cell types, as well as microglia, is evoked. ATP releasing from the dying and damaged cells acts as a DAMP to activate microglia for inflammatory IL-1β and IL-6 gene expression. High concentration of ATP might also promote RPE cell death under NaIO 3 stress.
    Figure Legend Snippet: Schematic representation of P2X7 activation in amplification of NaIO 3 -induced neuroinflammation and retinal cell death. ( Left ) In normal conditions, P2X7 is constitutively and highly expressed in microglia than photoreceptors, RPE and Müller cells. ( Right ) Upon NaIO 3 treatment, P2X7 expression in photoreceptors, Müller cells and RPE cells is increased, and cell death in these cell types, as well as microglia, is evoked. ATP releasing from the dying and damaged cells acts as a DAMP to activate microglia for inflammatory IL-1β and IL-6 gene expression. High concentration of ATP might also promote RPE cell death under NaIO 3 stress.

    Techniques Used: Activation Assay, Amplification, Expressing, Concentration Assay

    p2x 7 apr 008  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95

    Structured Review

    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, 2023-01
    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

    affinity purified polyclonal rabbit anti rat p2x 7 r  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Alomone Labs affinity purified polyclonal rabbit anti rat p2x 7 r
    Confocal fluorescence Z stack images of flat mount bladder mucosa taken from the urothelial towards the serosal surface. (A) Granular staining for Panx1 channels (red) is observed throughout the mucosa. Staining for the intermediate filament vimentin (green) is observed on the apical urothelial region and particularly on a few cells in the lamina propria, which likely correspond to suburothelial myofibroblasts. Note partial colocalization of Pannexin 1 with vimentin-positive cells. (B) Positive staining for <t>P2X</t> <t>7</t> <t>R</t> is observed in the urothelium, blood vessels (white arrows) and lamina propria, while staining for cytokeratin 7/17 is restricted to urothelial cells. Note intense P2X 7 R immunoreactivity on the basal region of the mucosa, which is likely localized to the lamina propria myofibroblasts. DAPI nuclear staining in blue. Scale bar = 20 µm.
    Affinity Purified Polyclonal Rabbit Anti Rat P2x 7 R, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/affinity purified polyclonal rabbit anti rat p2x 7 r/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    affinity purified polyclonal rabbit anti rat p2x 7 r - by Bioz Stars, 2023-01
    86/100 stars

    Images

    1) Product Images from "Pannexin 1 Channels Play Essential Roles in Urothelial Mechanotransduction and Intercellular Signaling"

    Article Title: Pannexin 1 Channels Play Essential Roles in Urothelial Mechanotransduction and Intercellular Signaling

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0106269

    Confocal fluorescence Z stack images of flat mount bladder mucosa taken from the urothelial towards the serosal surface. (A) Granular staining for Panx1 channels (red) is observed throughout the mucosa. Staining for the intermediate filament vimentin (green) is observed on the apical urothelial region and particularly on a few cells in the lamina propria, which likely correspond to suburothelial myofibroblasts. Note partial colocalization of Pannexin 1 with vimentin-positive cells. (B) Positive staining for P2X 7 R is observed in the urothelium, blood vessels (white arrows) and lamina propria, while staining for cytokeratin 7/17 is restricted to urothelial cells. Note intense P2X 7 R immunoreactivity on the basal region of the mucosa, which is likely localized to the lamina propria myofibroblasts. DAPI nuclear staining in blue. Scale bar = 20 µm.
    Figure Legend Snippet: Confocal fluorescence Z stack images of flat mount bladder mucosa taken from the urothelial towards the serosal surface. (A) Granular staining for Panx1 channels (red) is observed throughout the mucosa. Staining for the intermediate filament vimentin (green) is observed on the apical urothelial region and particularly on a few cells in the lamina propria, which likely correspond to suburothelial myofibroblasts. Note partial colocalization of Pannexin 1 with vimentin-positive cells. (B) Positive staining for P2X 7 R is observed in the urothelium, blood vessels (white arrows) and lamina propria, while staining for cytokeratin 7/17 is restricted to urothelial cells. Note intense P2X 7 R immunoreactivity on the basal region of the mucosa, which is likely localized to the lamina propria myofibroblasts. DAPI nuclear staining in blue. Scale bar = 20 µm.

    Techniques Used: Fluorescence, Staining

    Whole bladders isolated from wildtype (WT), Panx1 deficient (Panx1 −/− ) and P2X 7 R deficient (P2X 7 R −/− ) mice were bathed and instilled with PBS+glucose (1 g/L). A filling-voiding cycle was simulated by bladder instillation for 8 min at 1.5 mL/h flow rate followed by 5 min no flow, after which the bladder was emptied and ATP release in the bladder lumen was quantified. Data represent mean ± SEM (N = 9 WT, 4 Panx1 −/− and 7 P2X 7 R −/− bladders. Compared to WT: * P< 0.05 and ** P< 0.01 by Student’s t -test).
    Figure Legend Snippet: Whole bladders isolated from wildtype (WT), Panx1 deficient (Panx1 −/− ) and P2X 7 R deficient (P2X 7 R −/− ) mice were bathed and instilled with PBS+glucose (1 g/L). A filling-voiding cycle was simulated by bladder instillation for 8 min at 1.5 mL/h flow rate followed by 5 min no flow, after which the bladder was emptied and ATP release in the bladder lumen was quantified. Data represent mean ± SEM (N = 9 WT, 4 Panx1 −/− and 7 P2X 7 R −/− bladders. Compared to WT: * P< 0.05 and ** P< 0.01 by Student’s t -test).

    Techniques Used: Isolation

    Detection of Panx1 and P2X 7 R mRNA by PCR in (A) and protein by immunoblotting in (B). Total RNA from human bladder and HeLa cells were used as reference for the PCR analyses, and whole HeLa cell lysates was used as reference for immunoblotting.
    Figure Legend Snippet: Detection of Panx1 and P2X 7 R mRNA by PCR in (A) and protein by immunoblotting in (B). Total RNA from human bladder and HeLa cells were used as reference for the PCR analyses, and whole HeLa cell lysates was used as reference for immunoblotting.

    Techniques Used: Western Blot

    (A) TRT-HU1 cells: YoPro-1 uptake induced by cell swelling (hypoosmotic shock, black line) was significantly reduced in the presence of the Panx1 channel blocker mefloquine (MFQ 100 nM; yellow line), the P2X 7 R blocker A438079 (10 µM; red line) and when both Panx1 and P2X 7 R were blocked (blue line). Except for hypoosmotic with MFQ vs. A438079 and hypoosmotic with A438079+MFQ vs. isoosmotic, all other comparisons were significantly different at time 2800 sec ( P <0.01, N≥6) by two-way repeated measures ANOVA, followed by Tukey’s multiple comparison. (B) Primary mouse urothelial cells: YoPro-1 uptake induced by hypoosmotic shock was significantly lower in P2X 7 R −/− (red line) compared to wildtype (WT) urothelial cells (black line). Dye uptake by P2X 7 R −/− cells was abolished in the presence of the Panx1 channel blocker mefloquine (MFQ, 100 nM; green line) and was absent in Panx1 −/− urothelial cells (blue line). WT vs P2X 7 R −/− , P2X 7 R −/− +MFQ and Panx1 −/− ( P< 0.001, N = 4) by ANOVA followed by Tukey’s multiple comparison.
    Figure Legend Snippet: (A) TRT-HU1 cells: YoPro-1 uptake induced by cell swelling (hypoosmotic shock, black line) was significantly reduced in the presence of the Panx1 channel blocker mefloquine (MFQ 100 nM; yellow line), the P2X 7 R blocker A438079 (10 µM; red line) and when both Panx1 and P2X 7 R were blocked (blue line). Except for hypoosmotic with MFQ vs. A438079 and hypoosmotic with A438079+MFQ vs. isoosmotic, all other comparisons were significantly different at time 2800 sec ( P <0.01, N≥6) by two-way repeated measures ANOVA, followed by Tukey’s multiple comparison. (B) Primary mouse urothelial cells: YoPro-1 uptake induced by hypoosmotic shock was significantly lower in P2X 7 R −/− (red line) compared to wildtype (WT) urothelial cells (black line). Dye uptake by P2X 7 R −/− cells was abolished in the presence of the Panx1 channel blocker mefloquine (MFQ, 100 nM; green line) and was absent in Panx1 −/− urothelial cells (blue line). WT vs P2X 7 R −/− , P2X 7 R −/− +MFQ and Panx1 −/− ( P< 0.001, N = 4) by ANOVA followed by Tukey’s multiple comparison.

    Techniques Used:

    (A) Mechanical stimulation imposed by rinsing the cells with bathing solution induced ATP release from TRT-HU1 cells that was significantly higher than that from non-stimulated cells when measured in the presence or absence of the Panx1 channel blocker mefloquine (MFQ, 100 nM; n = 4 each, * P <0.05 and ** P <0.01, by paired t -test.). (B) Normalized ATP release with respect to basal values, however, was significantly lower in MFQ-treated compared to untreated cells (N = 4, $ P <0.05 by Mann-Whitney U test). (C) Exposure to low divalent cation solution (LDPBS), a condition known to enhance P2X 7 R activation, significantly increased ATP release from TRT-HU1 cells. All data represent mean ± SEM (N = 4 each, * P <0.05 by Student’s t -test).
    Figure Legend Snippet: (A) Mechanical stimulation imposed by rinsing the cells with bathing solution induced ATP release from TRT-HU1 cells that was significantly higher than that from non-stimulated cells when measured in the presence or absence of the Panx1 channel blocker mefloquine (MFQ, 100 nM; n = 4 each, * P <0.05 and ** P <0.01, by paired t -test.). (B) Normalized ATP release with respect to basal values, however, was significantly lower in MFQ-treated compared to untreated cells (N = 4, $ P <0.05 by Mann-Whitney U test). (C) Exposure to low divalent cation solution (LDPBS), a condition known to enhance P2X 7 R activation, significantly increased ATP release from TRT-HU1 cells. All data represent mean ± SEM (N = 4 each, * P <0.05 by Student’s t -test).

    Techniques Used: MANN-WHITNEY, Activation Assay

    anti p2x7r  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97

    Structured Review

    Alomone Labs anti p2x7r
    Primary antibodies used for Western Blot (WB) and immunohistochemistry (IHC).
    Anti P2x7r, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p2x7r/product/Alomone Labs
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti p2x7r - by Bioz Stars, 2023-01
    97/100 stars

    Images

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

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

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0117056

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

    Techniques Used: Western Blot, Immunohistochemistry

    p2x7r  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97

    Structured Review

    Alomone Labs p2x7r
    Sequences of siRNA to <t> P2X7R. </t>
    P2x7r, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p2x7r/product/Alomone Labs
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p2x7r - by Bioz Stars, 2023-01
    97/100 stars

    Images

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

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

    Journal: Oxidative Medicine and Cellular Longevity

    doi: 10.1155/2020/7956274

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

    Techniques Used:

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

    Techniques Used: Sequencing

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

    Techniques Used: Sequencing

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

    Techniques Used: Staining, Expressing

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

    Techniques Used: Immunofluorescence, Expressing, Western Blot

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

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

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

    Techniques Used: Activation Assay, Inhibition, Expressing

    p2x7 receptor c terminus epitope  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97

    Structured Review

    Alomone Labs p2x7 receptor c terminus epitope
    <t>P2X7</t> receptor and Oct-4 expression levels in undifferentiated (und) and cells induced to differentiation (days 0–8) were determined by ( A ) real-time PCR and ( B,C ) Western blotting assays as described in Materials and . For real-time PCR, quantitative analysis of the relative expression of P2X7R and Oct-4 in E14Tg2A cell line was performed using GAPDH mRNA transcription rates as endogenous control for normalization of expression levels. For Western blotting, P2X7R and Oct-4 expression levels were obtained and analyzed by densimetric analysis of protein bands and were compared to β-actin expression levels. The P2X7R was identified by two antibodies, which recognize the extracellular or C-terminus domain. Bars represent mean ± standard errors (S.E.) of three independent experiments performed in triplicate. Data were analyzed for statistical relevance with the One-Way ANOVA test followed by the Bonferroni post hoc test (*p<0,05,**p<0,01, ***p<0.001 compared to control data).
    P2x7 Receptor C Terminus Epitope, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p2x7 receptor c terminus epitope/product/Alomone Labs
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p2x7 receptor c terminus epitope - by Bioz Stars, 2023-01
    97/100 stars

    Images

    1) Product Images from "Modulation of Mouse Embryonic Stem Cell Proliferation and Neural Differentiation by the P2X7 Receptor"

    Article Title: Modulation of Mouse Embryonic Stem Cell Proliferation and Neural Differentiation by the P2X7 Receptor

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0096281

    P2X7 receptor and Oct-4 expression levels in undifferentiated (und) and cells induced to differentiation (days 0–8) were determined by ( A ) real-time PCR and ( B,C ) Western blotting assays as described in Materials and . For real-time PCR, quantitative analysis of the relative expression of P2X7R and Oct-4 in E14Tg2A cell line was performed using GAPDH mRNA transcription rates as endogenous control for normalization of expression levels. For Western blotting, P2X7R and Oct-4 expression levels were obtained and analyzed by densimetric analysis of protein bands and were compared to β-actin expression levels. The P2X7R was identified by two antibodies, which recognize the extracellular or C-terminus domain. Bars represent mean ± standard errors (S.E.) of three independent experiments performed in triplicate. Data were analyzed for statistical relevance with the One-Way ANOVA test followed by the Bonferroni post hoc test (*p<0,05,**p<0,01, ***p<0.001 compared to control data).
    Figure Legend Snippet: P2X7 receptor and Oct-4 expression levels in undifferentiated (und) and cells induced to differentiation (days 0–8) were determined by ( A ) real-time PCR and ( B,C ) Western blotting assays as described in Materials and . For real-time PCR, quantitative analysis of the relative expression of P2X7R and Oct-4 in E14Tg2A cell line was performed using GAPDH mRNA transcription rates as endogenous control for normalization of expression levels. For Western blotting, P2X7R and Oct-4 expression levels were obtained and analyzed by densimetric analysis of protein bands and were compared to β-actin expression levels. The P2X7R was identified by two antibodies, which recognize the extracellular or C-terminus domain. Bars represent mean ± standard errors (S.E.) of three independent experiments performed in triplicate. Data were analyzed for statistical relevance with the One-Way ANOVA test followed by the Bonferroni post hoc test (*p<0,05,**p<0,01, ***p<0.001 compared to control data).

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

    ( A ) P2X7 receptor isoform A, B, C and k expression in undifferentiated (UND) and cells following 8 days of neural differentiation (DIFF) was determined by RT-PCR. ( B ) P2X7R isoforms expression levels were obtained and analyzed by densimetric analysis of DNA bands and were compared to GAPDH expression levels. Data were analyzed for statistical relevance with the One-Way ANOVA test followed by the Bonferroni post hoc test (*p<0,05,**p<0,01, ***p<0.001 compared to control data).
    Figure Legend Snippet: ( A ) P2X7 receptor isoform A, B, C and k expression in undifferentiated (UND) and cells following 8 days of neural differentiation (DIFF) was determined by RT-PCR. ( B ) P2X7R isoforms expression levels were obtained and analyzed by densimetric analysis of DNA bands and were compared to GAPDH expression levels. Data were analyzed for statistical relevance with the One-Way ANOVA test followed by the Bonferroni post hoc test (*p<0,05,**p<0,01, ***p<0.001 compared to control data).

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

    anti purinergic p2x 7 r antibody  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    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, 2023-01
    94/100 stars

    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

    p2x7r  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97

    Structured Review

    Alomone Labs p2x7r
    Spleen cells from 3 to 4-mo-old MRL +/+ ( solid line ), MRL/ lpr ( dashed line ) and P2X7-deficient B6 <t>P2X7R−/−</t> ( dotted line ) mice (n = 3 mice/group) were treated for 45 min at 37°C with doses of ATP ranging from 100 to 5000 µM. Spleen cells were then triple-stained with anti-CD90, anti-CD19 and anti-CD62L mAb to assess by flow cytometry: (A) the percentage of CD62L-expressing CD19 – CD90 + T cells and (B) MFI of CD62L on CD19 – CD90 + T cells. Results are expressed as the mean percentage of initial expression ± SE.
    P2x7r, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p2x7r/product/Alomone Labs
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p2x7r - by Bioz Stars, 2023-01
    97/100 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

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 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.
    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, 2023-01
    95/100 stars
      Buy from Supplier

    86
    Alomone Labs rabbit anti p2x7
    Effect of Schisandrin B on expression levels of <t>P2X7</t> receptor in SCG of DM rats. (a) The schedule of experiment process. (b) The protein level of P2X7 protein in all group was measured by Western blotting. (c) The relative expression level of P2X7 protein to individual β -actin internal control was shown. Data are mean ± SD from four independent experiments. ( ∗ P < 0.05, ∗∗ P < 0.01 compared with the control group; # P < 0.05 compared with the DM group).
    Rabbit Anti P2x7, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti p2x7/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti p2x7 - by Bioz Stars, 2023-01
    86/100 stars
      Buy from Supplier

    86
    Alomone Labs p2x7 c term
    <t>P2X7</t> −/− exerted protection against NaIO 3 -induced photoreceptor injury. ( A ) Representative scotopic ERG responses from WT and P2X7 −/− mice with or without NaIO 3 (25 mg/kg) intraperitoneal injection. Three days after injection, ERG analysis was performed. ( B ) Quantification of the average amplitudes of a and b waves from WT control mice ( n = 8), WT NaIO 3 -treated mice ( n = 8), P2X7 −/− control mice ( n = 8) and P2X7 −/− NaIO 3 -treated mice ( n = 10). ( C ) Quantification of the implicit time of a and b waves from the 4 groups of mice. The data were presented as the mean ± SEM. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .
    P2x7 C Term, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p2x7 c term/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p2x7 c term - by Bioz Stars, 2023-01
    86/100 stars
      Buy from Supplier

    86
    Alomone Labs p2x7 extracellular ecto
    <t>P2X7</t> −/− exerted protection against NaIO 3 -induced photoreceptor injury. ( A ) Representative scotopic ERG responses from WT and P2X7 −/− mice with or without NaIO 3 (25 mg/kg) intraperitoneal injection. Three days after injection, ERG analysis was performed. ( B ) Quantification of the average amplitudes of a and b waves from WT control mice ( n = 8), WT NaIO 3 -treated mice ( n = 8), P2X7 −/− control mice ( n = 8) and P2X7 −/− NaIO 3 -treated mice ( n = 10). ( C ) Quantification of the implicit time of a and b waves from the 4 groups of mice. The data were presented as the mean ± SEM. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .
    P2x7 Extracellular Ecto, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p2x7 extracellular ecto/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p2x7 extracellular ecto - by Bioz Stars, 2023-01
    86/100 stars
      Buy from Supplier

    86
    Alomone Labs affinity purified polyclonal rabbit anti rat p2x 7 r
    Confocal fluorescence Z stack images of flat mount bladder mucosa taken from the urothelial towards the serosal surface. (A) Granular staining for Panx1 channels (red) is observed throughout the mucosa. Staining for the intermediate filament vimentin (green) is observed on the apical urothelial region and particularly on a few cells in the lamina propria, which likely correspond to suburothelial myofibroblasts. Note partial colocalization of Pannexin 1 with vimentin-positive cells. (B) Positive staining for <t>P2X</t> <t>7</t> <t>R</t> is observed in the urothelium, blood vessels (white arrows) and lamina propria, while staining for cytokeratin 7/17 is restricted to urothelial cells. Note intense P2X 7 R immunoreactivity on the basal region of the mucosa, which is likely localized to the lamina propria myofibroblasts. DAPI nuclear staining in blue. Scale bar = 20 µm.
    Affinity Purified Polyclonal Rabbit Anti Rat P2x 7 R, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/affinity purified polyclonal rabbit anti rat p2x 7 r/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    affinity purified polyclonal rabbit anti rat p2x 7 r - by Bioz Stars, 2023-01
    86/100 stars
      Buy from Supplier

    97
    Alomone Labs anti p2x7r
    Primary antibodies used for Western Blot (WB) and immunohistochemistry (IHC).
    Anti P2x7r, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p2x7r/product/Alomone Labs
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti p2x7r - by Bioz Stars, 2023-01
    97/100 stars
      Buy from Supplier

    97
    Alomone Labs p2x7r
    Sequences of siRNA to <t> P2X7R. </t>
    P2x7r, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p2x7r/product/Alomone Labs
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p2x7r - by Bioz Stars, 2023-01
    97/100 stars
      Buy from Supplier

    97
    Alomone Labs p2x7 receptor c terminus epitope
    <t>P2X7</t> receptor and Oct-4 expression levels in undifferentiated (und) and cells induced to differentiation (days 0–8) were determined by ( A ) real-time PCR and ( B,C ) Western blotting assays as described in Materials and . For real-time PCR, quantitative analysis of the relative expression of P2X7R and Oct-4 in E14Tg2A cell line was performed using GAPDH mRNA transcription rates as endogenous control for normalization of expression levels. For Western blotting, P2X7R and Oct-4 expression levels were obtained and analyzed by densimetric analysis of protein bands and were compared to β-actin expression levels. The P2X7R was identified by two antibodies, which recognize the extracellular or C-terminus domain. Bars represent mean ± standard errors (S.E.) of three independent experiments performed in triplicate. Data were analyzed for statistical relevance with the One-Way ANOVA test followed by the Bonferroni post hoc test (*p<0,05,**p<0,01, ***p<0.001 compared to control data).
    P2x7 Receptor C Terminus Epitope, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p2x7 receptor c terminus epitope/product/Alomone Labs
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p2x7 receptor c terminus epitope - by Bioz Stars, 2023-01
    97/100 stars
      Buy from Supplier

    94
    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, 2023-01
    94/100 stars
      Buy from Supplier

    Image Search Results


    (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

    Effect of Schisandrin B on expression levels of P2X7 receptor in SCG of DM rats. (a) The schedule of experiment process. (b) The protein level of P2X7 protein in all group was measured by Western blotting. (c) The relative expression level of P2X7 protein to individual β -actin internal control was shown. Data are mean ± SD from four independent experiments. ( ∗ P < 0.05, ∗∗ P < 0.01 compared with the control group; # P < 0.05 compared with the DM group).

    Journal: Disease Markers

    Article Title: Schisandrin B Alleviates Diabetic Cardiac Autonomic neuropathy Induced by P2X7 Receptor in Superior Cervical Ganglion via NLRP3

    doi: 10.1155/2023/9956950

    Figure Lengend Snippet: Effect of Schisandrin B on expression levels of P2X7 receptor in SCG of DM rats. (a) The schedule of experiment process. (b) The protein level of P2X7 protein in all group was measured by Western blotting. (c) The relative expression level of P2X7 protein to individual β -actin internal control was shown. Data are mean ± SD from four independent experiments. ( ∗ P < 0.05, ∗∗ P < 0.01 compared with the control group; # P < 0.05 compared with the DM group).

    Article Snippet: After 30-min incubation with goat serum blocking solution, the sections were incubated with mouse anti-GS (1 : 150, ab64613; Proteintech) and rabbit anti-P2X7 (1 : 150, APR-002; alomone labs) overnight at 4°C in cassette, followed by rinsing and incubating sections with TRITC-labeled goat anti-rabbit IgG (1 : 150, E031320-01; EARTHYX) or FITC-labeled goat anti-mouse IgG (1 : 150, E031210-01; EARTHYX) for 1 h at 37°C.

    Techniques: Expressing, Western Blot

    Effect of Schisandrin B on heart rate and blood pressure in rats.

    Journal: Disease Markers

    Article Title: Schisandrin B Alleviates Diabetic Cardiac Autonomic neuropathy Induced by P2X7 Receptor in Superior Cervical Ganglion via NLRP3

    doi: 10.1155/2023/9956950

    Figure Lengend Snippet: Effect of Schisandrin B on heart rate and blood pressure in rats.

    Article Snippet: After 30-min incubation with goat serum blocking solution, the sections were incubated with mouse anti-GS (1 : 150, ab64613; Proteintech) and rabbit anti-P2X7 (1 : 150, APR-002; alomone labs) overnight at 4°C in cassette, followed by rinsing and incubating sections with TRITC-labeled goat anti-rabbit IgG (1 : 150, E031320-01; EARTHYX) or FITC-labeled goat anti-mouse IgG (1 : 150, E031210-01; EARTHYX) for 1 h at 37°C.

    Techniques: shRNA

    Effects of Schisandrin B on heart rate variability in rats.

    Journal: Disease Markers

    Article Title: Schisandrin B Alleviates Diabetic Cardiac Autonomic neuropathy Induced by P2X7 Receptor in Superior Cervical Ganglion via NLRP3

    doi: 10.1155/2023/9956950

    Figure Lengend Snippet: Effects of Schisandrin B on heart rate variability in rats.

    Article Snippet: After 30-min incubation with goat serum blocking solution, the sections were incubated with mouse anti-GS (1 : 150, ab64613; Proteintech) and rabbit anti-P2X7 (1 : 150, APR-002; alomone labs) overnight at 4°C in cassette, followed by rinsing and incubating sections with TRITC-labeled goat anti-rabbit IgG (1 : 150, E031320-01; EARTHYX) or FITC-labeled goat anti-mouse IgG (1 : 150, E031210-01; EARTHYX) for 1 h at 37°C.

    Techniques: shRNA

    The double-label immunofluorescence staining of P2X7 and GS in SCG. The expression of P2X7 receptor and GS in SCG was detected by double-label immunofluorescence staining. The green signal represents GS staining with FITC, and the red signal indicates P2X7 staining with TRITC. The merged image represents the double staining of P2X7 and GS. Scale bar, 20 μ m.

    Journal: Disease Markers

    Article Title: Schisandrin B Alleviates Diabetic Cardiac Autonomic neuropathy Induced by P2X7 Receptor in Superior Cervical Ganglion via NLRP3

    doi: 10.1155/2023/9956950

    Figure Lengend Snippet: The double-label immunofluorescence staining of P2X7 and GS in SCG. The expression of P2X7 receptor and GS in SCG was detected by double-label immunofluorescence staining. The green signal represents GS staining with FITC, and the red signal indicates P2X7 staining with TRITC. The merged image represents the double staining of P2X7 and GS. Scale bar, 20 μ m.

    Article Snippet: After 30-min incubation with goat serum blocking solution, the sections were incubated with mouse anti-GS (1 : 150, ab64613; Proteintech) and rabbit anti-P2X7 (1 : 150, APR-002; alomone labs) overnight at 4°C in cassette, followed by rinsing and incubating sections with TRITC-labeled goat anti-rabbit IgG (1 : 150, E031320-01; EARTHYX) or FITC-labeled goat anti-mouse IgG (1 : 150, E031210-01; EARTHYX) for 1 h at 37°C.

    Techniques: Immunofluorescence, Staining, Expressing, Double Staining

    P2X7 −/− exerted protection against NaIO 3 -induced photoreceptor injury. ( A ) Representative scotopic ERG responses from WT and P2X7 −/− mice with or without NaIO 3 (25 mg/kg) intraperitoneal injection. Three days after injection, ERG analysis was performed. ( B ) Quantification of the average amplitudes of a and b waves from WT control mice ( n = 8), WT NaIO 3 -treated mice ( n = 8), P2X7 −/− control mice ( n = 8) and P2X7 −/− NaIO 3 -treated mice ( n = 10). ( C ) Quantification of the implicit time of a and b waves from the 4 groups of mice. The data were presented as the mean ± SEM. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Journal: Antioxidants

    Article Title: P2X7 Is Involved in the Mouse Retinal Degeneration via the Coordinated Actions in Different Retinal Cell Types

    doi: 10.3390/antiox12010141

    Figure Lengend Snippet: P2X7 −/− exerted protection against NaIO 3 -induced photoreceptor injury. ( A ) Representative scotopic ERG responses from WT and P2X7 −/− mice with or without NaIO 3 (25 mg/kg) intraperitoneal injection. Three days after injection, ERG analysis was performed. ( B ) Quantification of the average amplitudes of a and b waves from WT control mice ( n = 8), WT NaIO 3 -treated mice ( n = 8), P2X7 −/− control mice ( n = 8) and P2X7 −/− NaIO 3 -treated mice ( n = 10). ( C ) Quantification of the implicit time of a and b waves from the 4 groups of mice. The data were presented as the mean ± SEM. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Article Snippet: Different proteins were determined by immunoblot analysis using antibodies for P2X7 extracellular (ecto) (APR-008 Alomone, Jerusalem, Israel), P2X7 (c-term) (APR-004 Alomone, Jerusalem, Israel), β-actin (sc47778 Santa Cruz, CA, USA) and enhanced Chemiluminescence Reagent Plus (Perkin Elmer, Wellesley, MA, USA).

    Techniques: Injection

    P2X7 knockout protected mice from NaIO 3 -induced retinal epithelium injury. OCT images in posterior pole region ( A ) and optical nerve area ( B ) revealed retinal pigment epithelium degeneration after the mice were treated with 25 mg/kg of NaIO 3 for 3 days. RPE damage (yellow arrow in ( A , B ) was observed in the WT group but not in the P2X7 −/− group treated with NaIO 3 . Hyperreflective spots in both the vitreous region (blue arrow in ( B )) and the retina (orange arrow in ( B ) were shown. The ONL thickness was marked by the red double arrowheads in ( B ). The mice numbers were 8, 8, 8 and 10 for WT control, WT NaIO 3 , P2X7 −/− control and P2X7 −/− NaIO 3 groups, respectively. Scale bar: 200 μm. RPE, retinal pigment epithelium; OS, outer segment; IS, inner segment; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer.

    Journal: Antioxidants

    Article Title: P2X7 Is Involved in the Mouse Retinal Degeneration via the Coordinated Actions in Different Retinal Cell Types

    doi: 10.3390/antiox12010141

    Figure Lengend Snippet: P2X7 knockout protected mice from NaIO 3 -induced retinal epithelium injury. OCT images in posterior pole region ( A ) and optical nerve area ( B ) revealed retinal pigment epithelium degeneration after the mice were treated with 25 mg/kg of NaIO 3 for 3 days. RPE damage (yellow arrow in ( A , B ) was observed in the WT group but not in the P2X7 −/− group treated with NaIO 3 . Hyperreflective spots in both the vitreous region (blue arrow in ( B )) and the retina (orange arrow in ( B ) were shown. The ONL thickness was marked by the red double arrowheads in ( B ). The mice numbers were 8, 8, 8 and 10 for WT control, WT NaIO 3 , P2X7 −/− control and P2X7 −/− NaIO 3 groups, respectively. Scale bar: 200 μm. RPE, retinal pigment epithelium; OS, outer segment; IS, inner segment; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer.

    Article Snippet: Different proteins were determined by immunoblot analysis using antibodies for P2X7 extracellular (ecto) (APR-008 Alomone, Jerusalem, Israel), P2X7 (c-term) (APR-004 Alomone, Jerusalem, Israel), β-actin (sc47778 Santa Cruz, CA, USA) and enhanced Chemiluminescence Reagent Plus (Perkin Elmer, Wellesley, MA, USA).

    Techniques: Knock-Out

    Histological H&E examination of retinal injury caused by NaIO 3 was alleviated in P2X7 −/− mice. Retinal morphology and loss of photoreceptors and cell nuclei were evaluated by H&E by light microscopy at 3 days post-intraperitoneal injection of NaIO 3 (25 mg/kg) in WT and P2X7 −/− mice. Shown were representative light microscopic images of H&E staining in retinal sections. Enlargement of cell bodies in the ONL and INL, pigment granules along the RPE layer (black arrows) and ONL thinning were observed. Quantitative analysis of ONL thickness was shown. The mice number was 12 for each group. Scale bar: 200 μm. * p < 0.05, indicating significant decrease in ONL thickness by NaIO 3. # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Journal: Antioxidants

    Article Title: P2X7 Is Involved in the Mouse Retinal Degeneration via the Coordinated Actions in Different Retinal Cell Types

    doi: 10.3390/antiox12010141

    Figure Lengend Snippet: Histological H&E examination of retinal injury caused by NaIO 3 was alleviated in P2X7 −/− mice. Retinal morphology and loss of photoreceptors and cell nuclei were evaluated by H&E by light microscopy at 3 days post-intraperitoneal injection of NaIO 3 (25 mg/kg) in WT and P2X7 −/− mice. Shown were representative light microscopic images of H&E staining in retinal sections. Enlargement of cell bodies in the ONL and INL, pigment granules along the RPE layer (black arrows) and ONL thinning were observed. Quantitative analysis of ONL thickness was shown. The mice number was 12 for each group. Scale bar: 200 μm. * p < 0.05, indicating significant decrease in ONL thickness by NaIO 3. # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Article Snippet: Different proteins were determined by immunoblot analysis using antibodies for P2X7 extracellular (ecto) (APR-008 Alomone, Jerusalem, Israel), P2X7 (c-term) (APR-004 Alomone, Jerusalem, Israel), β-actin (sc47778 Santa Cruz, CA, USA) and enhanced Chemiluminescence Reagent Plus (Perkin Elmer, Wellesley, MA, USA).

    Techniques: Light Microscopy, Injection, Staining

    Histological examination of PAS and Masson trichrome staining. Representative light microscope images of PAS ( A ) and Masson trichrome ( B ) staining of retinal sections in WT and P2X7 −/− mice at 3 days post-intraperitoneal injection of NaIO 3 (25 mg/kg) were shown. The visible retinal pigment granules were shown as black arrows in both PAS ( A ) and Masson trichrome ( B ) staining. The mice number for PAS staining was 5 for each group and for Masson trichrome staining was 7 for each group. Scale bar: 200 μm.

    Journal: Antioxidants

    Article Title: P2X7 Is Involved in the Mouse Retinal Degeneration via the Coordinated Actions in Different Retinal Cell Types

    doi: 10.3390/antiox12010141

    Figure Lengend Snippet: Histological examination of PAS and Masson trichrome staining. Representative light microscope images of PAS ( A ) and Masson trichrome ( B ) staining of retinal sections in WT and P2X7 −/− mice at 3 days post-intraperitoneal injection of NaIO 3 (25 mg/kg) were shown. The visible retinal pigment granules were shown as black arrows in both PAS ( A ) and Masson trichrome ( B ) staining. The mice number for PAS staining was 5 for each group and for Masson trichrome staining was 7 for each group. Scale bar: 200 μm.

    Article Snippet: Different proteins were determined by immunoblot analysis using antibodies for P2X7 extracellular (ecto) (APR-008 Alomone, Jerusalem, Israel), P2X7 (c-term) (APR-004 Alomone, Jerusalem, Israel), β-actin (sc47778 Santa Cruz, CA, USA) and enhanced Chemiluminescence Reagent Plus (Perkin Elmer, Wellesley, MA, USA).

    Techniques: Staining, Light Microscopy, Injection

    NaIO 3 -induced neuroinflammation in retina was reversed by P2X7 knockout. After 36 h post-injection of NaIO 3 (25 mg/kg), mice retina was collected. ( A ) Total mRNA was extracted and reversely transcribed for quantitative PCR analysis of NLRP3, IL-1β and IL-6 mRNA expression. The mice number for qPCR analysis was 8 for each group. Values were normalized to β-actin gene expression and were expressed relative to the control group. ( B ) Parallelly, serum from mice was collected and subjected to ELISA to determine the amounts of IL-1β, TNF-α and IL-6, respectively. The mice number for ELISA was 5 for each group. The data were presented as the mean ± SEM. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Journal: Antioxidants

    Article Title: P2X7 Is Involved in the Mouse Retinal Degeneration via the Coordinated Actions in Different Retinal Cell Types

    doi: 10.3390/antiox12010141

    Figure Lengend Snippet: NaIO 3 -induced neuroinflammation in retina was reversed by P2X7 knockout. After 36 h post-injection of NaIO 3 (25 mg/kg), mice retina was collected. ( A ) Total mRNA was extracted and reversely transcribed for quantitative PCR analysis of NLRP3, IL-1β and IL-6 mRNA expression. The mice number for qPCR analysis was 8 for each group. Values were normalized to β-actin gene expression and were expressed relative to the control group. ( B ) Parallelly, serum from mice was collected and subjected to ELISA to determine the amounts of IL-1β, TNF-α and IL-6, respectively. The mice number for ELISA was 5 for each group. The data were presented as the mean ± SEM. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Article Snippet: Different proteins were determined by immunoblot analysis using antibodies for P2X7 extracellular (ecto) (APR-008 Alomone, Jerusalem, Israel), P2X7 (c-term) (APR-004 Alomone, Jerusalem, Israel), β-actin (sc47778 Santa Cruz, CA, USA) and enhanced Chemiluminescence Reagent Plus (Perkin Elmer, Wellesley, MA, USA).

    Techniques: Knock-Out, Injection, Real-time Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay

    NaIO 3 -induced caspase expression in retina was reversed by P2X7 knockout . ( A ) After 3 days post-injection of NaIO 3 (25 mg/kg), mice retina was collected, and total mRNA was extracted and reversely transcribed for quantitative PCR analyses of caspase-3, caspase-7 and caspase-8 mRNA expression. The mice number for qPCR analysis was 5 for each group. Values were normalized to β-actin gene expression and were expressed relative to the control group. ( B ) Using the retinal tissues to measure the cellular contents of total NADP, NADPH and NADP/NADPH as per manufacturer’s instructions. The mice number for NADPH activity was 5 for each group. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Journal: Antioxidants

    Article Title: P2X7 Is Involved in the Mouse Retinal Degeneration via the Coordinated Actions in Different Retinal Cell Types

    doi: 10.3390/antiox12010141

    Figure Lengend Snippet: NaIO 3 -induced caspase expression in retina was reversed by P2X7 knockout . ( A ) After 3 days post-injection of NaIO 3 (25 mg/kg), mice retina was collected, and total mRNA was extracted and reversely transcribed for quantitative PCR analyses of caspase-3, caspase-7 and caspase-8 mRNA expression. The mice number for qPCR analysis was 5 for each group. Values were normalized to β-actin gene expression and were expressed relative to the control group. ( B ) Using the retinal tissues to measure the cellular contents of total NADP, NADPH and NADP/NADPH as per manufacturer’s instructions. The mice number for NADPH activity was 5 for each group. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Article Snippet: Different proteins were determined by immunoblot analysis using antibodies for P2X7 extracellular (ecto) (APR-008 Alomone, Jerusalem, Israel), P2X7 (c-term) (APR-004 Alomone, Jerusalem, Israel), β-actin (sc47778 Santa Cruz, CA, USA) and enhanced Chemiluminescence Reagent Plus (Perkin Elmer, Wellesley, MA, USA).

    Techniques: Expressing, Knock-Out, Injection, Real-time Polymerase Chain Reaction, Activity Assay

    Comparison of P2X7 expression and effects of BzATP on cell viability in BV-2, 661W, ARPE-19 and rMC1 cells. ( A ) P2X7 protein level in each cell type was determined by immunoblotting. ( B , C ) Cells were treated with NaIO 3 (30 mM) for the indicated times. P2X7 mRNA level was determined by real-time PCR ( B ) and P2X7 protein was determined by immunoblotting ( C ). ( D ) Each cell type was treated with vehicle or BzATP (200 μM) for 6 or 18 h. Cell death was determined by Annexin V/PI staining with FACS. * p < 0.05, indicating the increased P2X7 gene expression by NaIO 3 ( B ) and the cell death caused by BzATP ( D ).

    Journal: Antioxidants

    Article Title: P2X7 Is Involved in the Mouse Retinal Degeneration via the Coordinated Actions in Different Retinal Cell Types

    doi: 10.3390/antiox12010141

    Figure Lengend Snippet: Comparison of P2X7 expression and effects of BzATP on cell viability in BV-2, 661W, ARPE-19 and rMC1 cells. ( A ) P2X7 protein level in each cell type was determined by immunoblotting. ( B , C ) Cells were treated with NaIO 3 (30 mM) for the indicated times. P2X7 mRNA level was determined by real-time PCR ( B ) and P2X7 protein was determined by immunoblotting ( C ). ( D ) Each cell type was treated with vehicle or BzATP (200 μM) for 6 or 18 h. Cell death was determined by Annexin V/PI staining with FACS. * p < 0.05, indicating the increased P2X7 gene expression by NaIO 3 ( B ) and the cell death caused by BzATP ( D ).

    Article Snippet: Different proteins were determined by immunoblot analysis using antibodies for P2X7 extracellular (ecto) (APR-008 Alomone, Jerusalem, Israel), P2X7 (c-term) (APR-004 Alomone, Jerusalem, Israel), β-actin (sc47778 Santa Cruz, CA, USA) and enhanced Chemiluminescence Reagent Plus (Perkin Elmer, Wellesley, MA, USA).

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

    Schematic representation of P2X7 activation in amplification of NaIO 3 -induced neuroinflammation and retinal cell death. ( Left ) In normal conditions, P2X7 is constitutively and highly expressed in microglia than photoreceptors, RPE and Müller cells. ( Right ) Upon NaIO 3 treatment, P2X7 expression in photoreceptors, Müller cells and RPE cells is increased, and cell death in these cell types, as well as microglia, is evoked. ATP releasing from the dying and damaged cells acts as a DAMP to activate microglia for inflammatory IL-1β and IL-6 gene expression. High concentration of ATP might also promote RPE cell death under NaIO 3 stress.

    Journal: Antioxidants

    Article Title: P2X7 Is Involved in the Mouse Retinal Degeneration via the Coordinated Actions in Different Retinal Cell Types

    doi: 10.3390/antiox12010141

    Figure Lengend Snippet: Schematic representation of P2X7 activation in amplification of NaIO 3 -induced neuroinflammation and retinal cell death. ( Left ) In normal conditions, P2X7 is constitutively and highly expressed in microglia than photoreceptors, RPE and Müller cells. ( Right ) Upon NaIO 3 treatment, P2X7 expression in photoreceptors, Müller cells and RPE cells is increased, and cell death in these cell types, as well as microglia, is evoked. ATP releasing from the dying and damaged cells acts as a DAMP to activate microglia for inflammatory IL-1β and IL-6 gene expression. High concentration of ATP might also promote RPE cell death under NaIO 3 stress.

    Article Snippet: Different proteins were determined by immunoblot analysis using antibodies for P2X7 extracellular (ecto) (APR-008 Alomone, Jerusalem, Israel), P2X7 (c-term) (APR-004 Alomone, Jerusalem, Israel), β-actin (sc47778 Santa Cruz, CA, USA) and enhanced Chemiluminescence Reagent Plus (Perkin Elmer, Wellesley, MA, USA).

    Techniques: Activation Assay, Amplification, Expressing, Concentration Assay

    P2X7 −/− exerted protection against NaIO 3 -induced photoreceptor injury. ( A ) Representative scotopic ERG responses from WT and P2X7 −/− mice with or without NaIO 3 (25 mg/kg) intraperitoneal injection. Three days after injection, ERG analysis was performed. ( B ) Quantification of the average amplitudes of a and b waves from WT control mice ( n = 8), WT NaIO 3 -treated mice ( n = 8), P2X7 −/− control mice ( n = 8) and P2X7 −/− NaIO 3 -treated mice ( n = 10). ( C ) Quantification of the implicit time of a and b waves from the 4 groups of mice. The data were presented as the mean ± SEM. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Journal: Antioxidants

    Article Title: P2X7 Is Involved in the Mouse Retinal Degeneration via the Coordinated Actions in Different Retinal Cell Types

    doi: 10.3390/antiox12010141

    Figure Lengend Snippet: P2X7 −/− exerted protection against NaIO 3 -induced photoreceptor injury. ( A ) Representative scotopic ERG responses from WT and P2X7 −/− mice with or without NaIO 3 (25 mg/kg) intraperitoneal injection. Three days after injection, ERG analysis was performed. ( B ) Quantification of the average amplitudes of a and b waves from WT control mice ( n = 8), WT NaIO 3 -treated mice ( n = 8), P2X7 −/− control mice ( n = 8) and P2X7 −/− NaIO 3 -treated mice ( n = 10). ( C ) Quantification of the implicit time of a and b waves from the 4 groups of mice. The data were presented as the mean ± SEM. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Article Snippet: Different proteins were determined by immunoblot analysis using antibodies for P2X7 extracellular (ecto) (APR-008 Alomone, Jerusalem, Israel), P2X7 (c-term) (APR-004 Alomone, Jerusalem, Israel), β-actin (sc47778 Santa Cruz, CA, USA) and enhanced Chemiluminescence Reagent Plus (Perkin Elmer, Wellesley, MA, USA).

    Techniques: Injection

    P2X7 knockout protected mice from NaIO 3 -induced retinal epithelium injury. OCT images in posterior pole region ( A ) and optical nerve area ( B ) revealed retinal pigment epithelium degeneration after the mice were treated with 25 mg/kg of NaIO 3 for 3 days. RPE damage (yellow arrow in ( A , B ) was observed in the WT group but not in the P2X7 −/− group treated with NaIO 3 . Hyperreflective spots in both the vitreous region (blue arrow in ( B )) and the retina (orange arrow in ( B ) were shown. The ONL thickness was marked by the red double arrowheads in ( B ). The mice numbers were 8, 8, 8 and 10 for WT control, WT NaIO 3 , P2X7 −/− control and P2X7 −/− NaIO 3 groups, respectively. Scale bar: 200 μm. RPE, retinal pigment epithelium; OS, outer segment; IS, inner segment; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer.

    Journal: Antioxidants

    Article Title: P2X7 Is Involved in the Mouse Retinal Degeneration via the Coordinated Actions in Different Retinal Cell Types

    doi: 10.3390/antiox12010141

    Figure Lengend Snippet: P2X7 knockout protected mice from NaIO 3 -induced retinal epithelium injury. OCT images in posterior pole region ( A ) and optical nerve area ( B ) revealed retinal pigment epithelium degeneration after the mice were treated with 25 mg/kg of NaIO 3 for 3 days. RPE damage (yellow arrow in ( A , B ) was observed in the WT group but not in the P2X7 −/− group treated with NaIO 3 . Hyperreflective spots in both the vitreous region (blue arrow in ( B )) and the retina (orange arrow in ( B ) were shown. The ONL thickness was marked by the red double arrowheads in ( B ). The mice numbers were 8, 8, 8 and 10 for WT control, WT NaIO 3 , P2X7 −/− control and P2X7 −/− NaIO 3 groups, respectively. Scale bar: 200 μm. RPE, retinal pigment epithelium; OS, outer segment; IS, inner segment; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer.

    Article Snippet: Different proteins were determined by immunoblot analysis using antibodies for P2X7 extracellular (ecto) (APR-008 Alomone, Jerusalem, Israel), P2X7 (c-term) (APR-004 Alomone, Jerusalem, Israel), β-actin (sc47778 Santa Cruz, CA, USA) and enhanced Chemiluminescence Reagent Plus (Perkin Elmer, Wellesley, MA, USA).

    Techniques: Knock-Out

    Histological H&E examination of retinal injury caused by NaIO 3 was alleviated in P2X7 −/− mice. Retinal morphology and loss of photoreceptors and cell nuclei were evaluated by H&E by light microscopy at 3 days post-intraperitoneal injection of NaIO 3 (25 mg/kg) in WT and P2X7 −/− mice. Shown were representative light microscopic images of H&E staining in retinal sections. Enlargement of cell bodies in the ONL and INL, pigment granules along the RPE layer (black arrows) and ONL thinning were observed. Quantitative analysis of ONL thickness was shown. The mice number was 12 for each group. Scale bar: 200 μm. * p < 0.05, indicating significant decrease in ONL thickness by NaIO 3. # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Journal: Antioxidants

    Article Title: P2X7 Is Involved in the Mouse Retinal Degeneration via the Coordinated Actions in Different Retinal Cell Types

    doi: 10.3390/antiox12010141

    Figure Lengend Snippet: Histological H&E examination of retinal injury caused by NaIO 3 was alleviated in P2X7 −/− mice. Retinal morphology and loss of photoreceptors and cell nuclei were evaluated by H&E by light microscopy at 3 days post-intraperitoneal injection of NaIO 3 (25 mg/kg) in WT and P2X7 −/− mice. Shown were representative light microscopic images of H&E staining in retinal sections. Enlargement of cell bodies in the ONL and INL, pigment granules along the RPE layer (black arrows) and ONL thinning were observed. Quantitative analysis of ONL thickness was shown. The mice number was 12 for each group. Scale bar: 200 μm. * p < 0.05, indicating significant decrease in ONL thickness by NaIO 3. # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Article Snippet: Different proteins were determined by immunoblot analysis using antibodies for P2X7 extracellular (ecto) (APR-008 Alomone, Jerusalem, Israel), P2X7 (c-term) (APR-004 Alomone, Jerusalem, Israel), β-actin (sc47778 Santa Cruz, CA, USA) and enhanced Chemiluminescence Reagent Plus (Perkin Elmer, Wellesley, MA, USA).

    Techniques: Light Microscopy, Injection, Staining

    Histological examination of PAS and Masson trichrome staining. Representative light microscope images of PAS ( A ) and Masson trichrome ( B ) staining of retinal sections in WT and P2X7 −/− mice at 3 days post-intraperitoneal injection of NaIO 3 (25 mg/kg) were shown. The visible retinal pigment granules were shown as black arrows in both PAS ( A ) and Masson trichrome ( B ) staining. The mice number for PAS staining was 5 for each group and for Masson trichrome staining was 7 for each group. Scale bar: 200 μm.

    Journal: Antioxidants

    Article Title: P2X7 Is Involved in the Mouse Retinal Degeneration via the Coordinated Actions in Different Retinal Cell Types

    doi: 10.3390/antiox12010141

    Figure Lengend Snippet: Histological examination of PAS and Masson trichrome staining. Representative light microscope images of PAS ( A ) and Masson trichrome ( B ) staining of retinal sections in WT and P2X7 −/− mice at 3 days post-intraperitoneal injection of NaIO 3 (25 mg/kg) were shown. The visible retinal pigment granules were shown as black arrows in both PAS ( A ) and Masson trichrome ( B ) staining. The mice number for PAS staining was 5 for each group and for Masson trichrome staining was 7 for each group. Scale bar: 200 μm.

    Article Snippet: Different proteins were determined by immunoblot analysis using antibodies for P2X7 extracellular (ecto) (APR-008 Alomone, Jerusalem, Israel), P2X7 (c-term) (APR-004 Alomone, Jerusalem, Israel), β-actin (sc47778 Santa Cruz, CA, USA) and enhanced Chemiluminescence Reagent Plus (Perkin Elmer, Wellesley, MA, USA).

    Techniques: Staining, Light Microscopy, Injection

    NaIO 3 -induced neuroinflammation in retina was reversed by P2X7 knockout. After 36 h post-injection of NaIO 3 (25 mg/kg), mice retina was collected. ( A ) Total mRNA was extracted and reversely transcribed for quantitative PCR analysis of NLRP3, IL-1β and IL-6 mRNA expression. The mice number for qPCR analysis was 8 for each group. Values were normalized to β-actin gene expression and were expressed relative to the control group. ( B ) Parallelly, serum from mice was collected and subjected to ELISA to determine the amounts of IL-1β, TNF-α and IL-6, respectively. The mice number for ELISA was 5 for each group. The data were presented as the mean ± SEM. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Journal: Antioxidants

    Article Title: P2X7 Is Involved in the Mouse Retinal Degeneration via the Coordinated Actions in Different Retinal Cell Types

    doi: 10.3390/antiox12010141

    Figure Lengend Snippet: NaIO 3 -induced neuroinflammation in retina was reversed by P2X7 knockout. After 36 h post-injection of NaIO 3 (25 mg/kg), mice retina was collected. ( A ) Total mRNA was extracted and reversely transcribed for quantitative PCR analysis of NLRP3, IL-1β and IL-6 mRNA expression. The mice number for qPCR analysis was 8 for each group. Values were normalized to β-actin gene expression and were expressed relative to the control group. ( B ) Parallelly, serum from mice was collected and subjected to ELISA to determine the amounts of IL-1β, TNF-α and IL-6, respectively. The mice number for ELISA was 5 for each group. The data were presented as the mean ± SEM. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Article Snippet: Different proteins were determined by immunoblot analysis using antibodies for P2X7 extracellular (ecto) (APR-008 Alomone, Jerusalem, Israel), P2X7 (c-term) (APR-004 Alomone, Jerusalem, Israel), β-actin (sc47778 Santa Cruz, CA, USA) and enhanced Chemiluminescence Reagent Plus (Perkin Elmer, Wellesley, MA, USA).

    Techniques: Knock-Out, Injection, Real-time Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay

    NaIO 3 -induced caspase expression in retina was reversed by P2X7 knockout . ( A ) After 3 days post-injection of NaIO 3 (25 mg/kg), mice retina was collected, and total mRNA was extracted and reversely transcribed for quantitative PCR analyses of caspase-3, caspase-7 and caspase-8 mRNA expression. The mice number for qPCR analysis was 5 for each group. Values were normalized to β-actin gene expression and were expressed relative to the control group. ( B ) Using the retinal tissues to measure the cellular contents of total NADP, NADPH and NADP/NADPH as per manufacturer’s instructions. The mice number for NADPH activity was 5 for each group. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Journal: Antioxidants

    Article Title: P2X7 Is Involved in the Mouse Retinal Degeneration via the Coordinated Actions in Different Retinal Cell Types

    doi: 10.3390/antiox12010141

    Figure Lengend Snippet: NaIO 3 -induced caspase expression in retina was reversed by P2X7 knockout . ( A ) After 3 days post-injection of NaIO 3 (25 mg/kg), mice retina was collected, and total mRNA was extracted and reversely transcribed for quantitative PCR analyses of caspase-3, caspase-7 and caspase-8 mRNA expression. The mice number for qPCR analysis was 5 for each group. Values were normalized to β-actin gene expression and were expressed relative to the control group. ( B ) Using the retinal tissues to measure the cellular contents of total NADP, NADPH and NADP/NADPH as per manufacturer’s instructions. The mice number for NADPH activity was 5 for each group. * p < 0.05 compared with the WT control group, # p < 0.05 compared with the WT control group treated with NaIO 3 .

    Article Snippet: Different proteins were determined by immunoblot analysis using antibodies for P2X7 extracellular (ecto) (APR-008 Alomone, Jerusalem, Israel), P2X7 (c-term) (APR-004 Alomone, Jerusalem, Israel), β-actin (sc47778 Santa Cruz, CA, USA) and enhanced Chemiluminescence Reagent Plus (Perkin Elmer, Wellesley, MA, USA).

    Techniques: Expressing, Knock-Out, Injection, Real-time Polymerase Chain Reaction, Activity Assay

    Comparison of P2X7 expression and effects of BzATP on cell viability in BV-2, 661W, ARPE-19 and rMC1 cells. ( A ) P2X7 protein level in each cell type was determined by immunoblotting. ( B , C ) Cells were treated with NaIO 3 (30 mM) for the indicated times. P2X7 mRNA level was determined by real-time PCR ( B ) and P2X7 protein was determined by immunoblotting ( C ). ( D ) Each cell type was treated with vehicle or BzATP (200 μM) for 6 or 18 h. Cell death was determined by Annexin V/PI staining with FACS. * p < 0.05, indicating the increased P2X7 gene expression by NaIO 3 ( B ) and the cell death caused by BzATP ( D ).

    Journal: Antioxidants

    Article Title: P2X7 Is Involved in the Mouse Retinal Degeneration via the Coordinated Actions in Different Retinal Cell Types

    doi: 10.3390/antiox12010141

    Figure Lengend Snippet: Comparison of P2X7 expression and effects of BzATP on cell viability in BV-2, 661W, ARPE-19 and rMC1 cells. ( A ) P2X7 protein level in each cell type was determined by immunoblotting. ( B , C ) Cells were treated with NaIO 3 (30 mM) for the indicated times. P2X7 mRNA level was determined by real-time PCR ( B ) and P2X7 protein was determined by immunoblotting ( C ). ( D ) Each cell type was treated with vehicle or BzATP (200 μM) for 6 or 18 h. Cell death was determined by Annexin V/PI staining with FACS. * p < 0.05, indicating the increased P2X7 gene expression by NaIO 3 ( B ) and the cell death caused by BzATP ( D ).

    Article Snippet: Different proteins were determined by immunoblot analysis using antibodies for P2X7 extracellular (ecto) (APR-008 Alomone, Jerusalem, Israel), P2X7 (c-term) (APR-004 Alomone, Jerusalem, Israel), β-actin (sc47778 Santa Cruz, CA, USA) and enhanced Chemiluminescence Reagent Plus (Perkin Elmer, Wellesley, MA, USA).

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

    Schematic representation of P2X7 activation in amplification of NaIO 3 -induced neuroinflammation and retinal cell death. ( Left ) In normal conditions, P2X7 is constitutively and highly expressed in microglia than photoreceptors, RPE and Müller cells. ( Right ) Upon NaIO 3 treatment, P2X7 expression in photoreceptors, Müller cells and RPE cells is increased, and cell death in these cell types, as well as microglia, is evoked. ATP releasing from the dying and damaged cells acts as a DAMP to activate microglia for inflammatory IL-1β and IL-6 gene expression. High concentration of ATP might also promote RPE cell death under NaIO 3 stress.

    Journal: Antioxidants

    Article Title: P2X7 Is Involved in the Mouse Retinal Degeneration via the Coordinated Actions in Different Retinal Cell Types

    doi: 10.3390/antiox12010141

    Figure Lengend Snippet: Schematic representation of P2X7 activation in amplification of NaIO 3 -induced neuroinflammation and retinal cell death. ( Left ) In normal conditions, P2X7 is constitutively and highly expressed in microglia than photoreceptors, RPE and Müller cells. ( Right ) Upon NaIO 3 treatment, P2X7 expression in photoreceptors, Müller cells and RPE cells is increased, and cell death in these cell types, as well as microglia, is evoked. ATP releasing from the dying and damaged cells acts as a DAMP to activate microglia for inflammatory IL-1β and IL-6 gene expression. High concentration of ATP might also promote RPE cell death under NaIO 3 stress.

    Article Snippet: Different proteins were determined by immunoblot analysis using antibodies for P2X7 extracellular (ecto) (APR-008 Alomone, Jerusalem, Israel), P2X7 (c-term) (APR-004 Alomone, Jerusalem, Israel), β-actin (sc47778 Santa Cruz, CA, USA) and enhanced Chemiluminescence Reagent Plus (Perkin Elmer, Wellesley, MA, USA).

    Techniques: Activation Assay, Amplification, Expressing, Concentration Assay

    Confocal fluorescence Z stack images of flat mount bladder mucosa taken from the urothelial towards the serosal surface. (A) Granular staining for Panx1 channels (red) is observed throughout the mucosa. Staining for the intermediate filament vimentin (green) is observed on the apical urothelial region and particularly on a few cells in the lamina propria, which likely correspond to suburothelial myofibroblasts. Note partial colocalization of Pannexin 1 with vimentin-positive cells. (B) Positive staining for P2X 7 R is observed in the urothelium, blood vessels (white arrows) and lamina propria, while staining for cytokeratin 7/17 is restricted to urothelial cells. Note intense P2X 7 R immunoreactivity on the basal region of the mucosa, which is likely localized to the lamina propria myofibroblasts. DAPI nuclear staining in blue. Scale bar = 20 µm.

    Journal: PLoS ONE

    Article Title: Pannexin 1 Channels Play Essential Roles in Urothelial Mechanotransduction and Intercellular Signaling

    doi: 10.1371/journal.pone.0106269

    Figure Lengend Snippet: Confocal fluorescence Z stack images of flat mount bladder mucosa taken from the urothelial towards the serosal surface. (A) Granular staining for Panx1 channels (red) is observed throughout the mucosa. Staining for the intermediate filament vimentin (green) is observed on the apical urothelial region and particularly on a few cells in the lamina propria, which likely correspond to suburothelial myofibroblasts. Note partial colocalization of Pannexin 1 with vimentin-positive cells. (B) Positive staining for P2X 7 R is observed in the urothelium, blood vessels (white arrows) and lamina propria, while staining for cytokeratin 7/17 is restricted to urothelial cells. Note intense P2X 7 R immunoreactivity on the basal region of the mucosa, which is likely localized to the lamina propria myofibroblasts. DAPI nuclear staining in blue. Scale bar = 20 µm.

    Article Snippet: The following primary antibodies were used: affinity purified polyclonal rabbit anti-rat P2X 7 R corresponding to amino acid residues 576–595 (1∶250, Alomone Labs, Jerusalem, Israel), polyclonal rabbit anti-mouse Pannexin 1 CL (Cytoplasmic loop, 1∶50, Invitrogen, Carlsbad, CA), monoclonal mouse anti-human cytokeratin 7/17 (1∶100, Santa Cruz Biotechnology, Dallas, TX) and monoclonal mouse anti-vimentin (1∶100, Sigma-Aldrich).

    Techniques: Fluorescence, Staining

    Whole bladders isolated from wildtype (WT), Panx1 deficient (Panx1 −/− ) and P2X 7 R deficient (P2X 7 R −/− ) mice were bathed and instilled with PBS+glucose (1 g/L). A filling-voiding cycle was simulated by bladder instillation for 8 min at 1.5 mL/h flow rate followed by 5 min no flow, after which the bladder was emptied and ATP release in the bladder lumen was quantified. Data represent mean ± SEM (N = 9 WT, 4 Panx1 −/− and 7 P2X 7 R −/− bladders. Compared to WT: * P< 0.05 and ** P< 0.01 by Student’s t -test).

    Journal: PLoS ONE

    Article Title: Pannexin 1 Channels Play Essential Roles in Urothelial Mechanotransduction and Intercellular Signaling

    doi: 10.1371/journal.pone.0106269

    Figure Lengend Snippet: Whole bladders isolated from wildtype (WT), Panx1 deficient (Panx1 −/− ) and P2X 7 R deficient (P2X 7 R −/− ) mice were bathed and instilled with PBS+glucose (1 g/L). A filling-voiding cycle was simulated by bladder instillation for 8 min at 1.5 mL/h flow rate followed by 5 min no flow, after which the bladder was emptied and ATP release in the bladder lumen was quantified. Data represent mean ± SEM (N = 9 WT, 4 Panx1 −/− and 7 P2X 7 R −/− bladders. Compared to WT: * P< 0.05 and ** P< 0.01 by Student’s t -test).

    Article Snippet: The following primary antibodies were used: affinity purified polyclonal rabbit anti-rat P2X 7 R corresponding to amino acid residues 576–595 (1∶250, Alomone Labs, Jerusalem, Israel), polyclonal rabbit anti-mouse Pannexin 1 CL (Cytoplasmic loop, 1∶50, Invitrogen, Carlsbad, CA), monoclonal mouse anti-human cytokeratin 7/17 (1∶100, Santa Cruz Biotechnology, Dallas, TX) and monoclonal mouse anti-vimentin (1∶100, Sigma-Aldrich).

    Techniques: Isolation

    Detection of Panx1 and P2X 7 R mRNA by PCR in (A) and protein by immunoblotting in (B). Total RNA from human bladder and HeLa cells were used as reference for the PCR analyses, and whole HeLa cell lysates was used as reference for immunoblotting.

    Journal: PLoS ONE

    Article Title: Pannexin 1 Channels Play Essential Roles in Urothelial Mechanotransduction and Intercellular Signaling

    doi: 10.1371/journal.pone.0106269

    Figure Lengend Snippet: Detection of Panx1 and P2X 7 R mRNA by PCR in (A) and protein by immunoblotting in (B). Total RNA from human bladder and HeLa cells were used as reference for the PCR analyses, and whole HeLa cell lysates was used as reference for immunoblotting.

    Article Snippet: The following primary antibodies were used: affinity purified polyclonal rabbit anti-rat P2X 7 R corresponding to amino acid residues 576–595 (1∶250, Alomone Labs, Jerusalem, Israel), polyclonal rabbit anti-mouse Pannexin 1 CL (Cytoplasmic loop, 1∶50, Invitrogen, Carlsbad, CA), monoclonal mouse anti-human cytokeratin 7/17 (1∶100, Santa Cruz Biotechnology, Dallas, TX) and monoclonal mouse anti-vimentin (1∶100, Sigma-Aldrich).

    Techniques: Western Blot

    (A) TRT-HU1 cells: YoPro-1 uptake induced by cell swelling (hypoosmotic shock, black line) was significantly reduced in the presence of the Panx1 channel blocker mefloquine (MFQ 100 nM; yellow line), the P2X 7 R blocker A438079 (10 µM; red line) and when both Panx1 and P2X 7 R were blocked (blue line). Except for hypoosmotic with MFQ vs. A438079 and hypoosmotic with A438079+MFQ vs. isoosmotic, all other comparisons were significantly different at time 2800 sec ( P <0.01, N≥6) by two-way repeated measures ANOVA, followed by Tukey’s multiple comparison. (B) Primary mouse urothelial cells: YoPro-1 uptake induced by hypoosmotic shock was significantly lower in P2X 7 R −/− (red line) compared to wildtype (WT) urothelial cells (black line). Dye uptake by P2X 7 R −/− cells was abolished in the presence of the Panx1 channel blocker mefloquine (MFQ, 100 nM; green line) and was absent in Panx1 −/− urothelial cells (blue line). WT vs P2X 7 R −/− , P2X 7 R −/− +MFQ and Panx1 −/− ( P< 0.001, N = 4) by ANOVA followed by Tukey’s multiple comparison.

    Journal: PLoS ONE

    Article Title: Pannexin 1 Channels Play Essential Roles in Urothelial Mechanotransduction and Intercellular Signaling

    doi: 10.1371/journal.pone.0106269

    Figure Lengend Snippet: (A) TRT-HU1 cells: YoPro-1 uptake induced by cell swelling (hypoosmotic shock, black line) was significantly reduced in the presence of the Panx1 channel blocker mefloquine (MFQ 100 nM; yellow line), the P2X 7 R blocker A438079 (10 µM; red line) and when both Panx1 and P2X 7 R were blocked (blue line). Except for hypoosmotic with MFQ vs. A438079 and hypoosmotic with A438079+MFQ vs. isoosmotic, all other comparisons were significantly different at time 2800 sec ( P <0.01, N≥6) by two-way repeated measures ANOVA, followed by Tukey’s multiple comparison. (B) Primary mouse urothelial cells: YoPro-1 uptake induced by hypoosmotic shock was significantly lower in P2X 7 R −/− (red line) compared to wildtype (WT) urothelial cells (black line). Dye uptake by P2X 7 R −/− cells was abolished in the presence of the Panx1 channel blocker mefloquine (MFQ, 100 nM; green line) and was absent in Panx1 −/− urothelial cells (blue line). WT vs P2X 7 R −/− , P2X 7 R −/− +MFQ and Panx1 −/− ( P< 0.001, N = 4) by ANOVA followed by Tukey’s multiple comparison.

    Article Snippet: The following primary antibodies were used: affinity purified polyclonal rabbit anti-rat P2X 7 R corresponding to amino acid residues 576–595 (1∶250, Alomone Labs, Jerusalem, Israel), polyclonal rabbit anti-mouse Pannexin 1 CL (Cytoplasmic loop, 1∶50, Invitrogen, Carlsbad, CA), monoclonal mouse anti-human cytokeratin 7/17 (1∶100, Santa Cruz Biotechnology, Dallas, TX) and monoclonal mouse anti-vimentin (1∶100, Sigma-Aldrich).

    Techniques:

    (A) Mechanical stimulation imposed by rinsing the cells with bathing solution induced ATP release from TRT-HU1 cells that was significantly higher than that from non-stimulated cells when measured in the presence or absence of the Panx1 channel blocker mefloquine (MFQ, 100 nM; n = 4 each, * P <0.05 and ** P <0.01, by paired t -test.). (B) Normalized ATP release with respect to basal values, however, was significantly lower in MFQ-treated compared to untreated cells (N = 4, $ P <0.05 by Mann-Whitney U test). (C) Exposure to low divalent cation solution (LDPBS), a condition known to enhance P2X 7 R activation, significantly increased ATP release from TRT-HU1 cells. All data represent mean ± SEM (N = 4 each, * P <0.05 by Student’s t -test).

    Journal: PLoS ONE

    Article Title: Pannexin 1 Channels Play Essential Roles in Urothelial Mechanotransduction and Intercellular Signaling

    doi: 10.1371/journal.pone.0106269

    Figure Lengend Snippet: (A) Mechanical stimulation imposed by rinsing the cells with bathing solution induced ATP release from TRT-HU1 cells that was significantly higher than that from non-stimulated cells when measured in the presence or absence of the Panx1 channel blocker mefloquine (MFQ, 100 nM; n = 4 each, * P <0.05 and ** P <0.01, by paired t -test.). (B) Normalized ATP release with respect to basal values, however, was significantly lower in MFQ-treated compared to untreated cells (N = 4, $ P <0.05 by Mann-Whitney U test). (C) Exposure to low divalent cation solution (LDPBS), a condition known to enhance P2X 7 R activation, significantly increased ATP release from TRT-HU1 cells. All data represent mean ± SEM (N = 4 each, * P <0.05 by Student’s t -test).

    Article Snippet: The following primary antibodies were used: affinity purified polyclonal rabbit anti-rat P2X 7 R corresponding to amino acid residues 576–595 (1∶250, Alomone Labs, Jerusalem, Israel), polyclonal rabbit anti-mouse Pannexin 1 CL (Cytoplasmic loop, 1∶50, Invitrogen, Carlsbad, CA), monoclonal mouse anti-human cytokeratin 7/17 (1∶100, Santa Cruz Biotechnology, Dallas, TX) and monoclonal mouse anti-vimentin (1∶100, Sigma-Aldrich).

    Techniques: MANN-WHITNEY, Activation Assay

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

    Journal: PLoS ONE

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

    doi: 10.1371/journal.pone.0117056

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

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

    Techniques: Western Blot, Immunohistochemistry

    Sequences of siRNA to  P2X7R.

    Journal: Oxidative Medicine and Cellular Longevity

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

    doi: 10.1155/2020/7956274

    Figure Lengend Snippet: Sequences of siRNA to P2X7R.

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

    Techniques:

    Rat RT-qPCR primer sequences.

    Journal: Oxidative Medicine and Cellular Longevity

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

    doi: 10.1155/2020/7956274

    Figure Lengend Snippet: Rat RT-qPCR primer sequences.

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

    Techniques: Sequencing

    Mice RT-qPCR primer sequences.

    Journal: Oxidative Medicine and Cellular Longevity

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

    doi: 10.1155/2020/7956274

    Figure Lengend Snippet: Mice RT-qPCR primer sequences.

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

    Techniques: Sequencing

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

    Journal: Oxidative Medicine and Cellular Longevity

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

    doi: 10.1155/2020/7956274

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

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

    Techniques: Staining, Expressing

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

    Journal: Oxidative Medicine and Cellular Longevity

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

    doi: 10.1155/2020/7956274

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

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

    Techniques: Immunofluorescence, Expressing, Western Blot

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

    Journal: Oxidative Medicine and Cellular Longevity

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

    doi: 10.1155/2020/7956274

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

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

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

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

    Journal: Oxidative Medicine and Cellular Longevity

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

    doi: 10.1155/2020/7956274

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

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

    Techniques: Activation Assay, Inhibition, Expressing

    P2X7 receptor and Oct-4 expression levels in undifferentiated (und) and cells induced to differentiation (days 0–8) were determined by ( A ) real-time PCR and ( B,C ) Western blotting assays as described in Materials and . For real-time PCR, quantitative analysis of the relative expression of P2X7R and Oct-4 in E14Tg2A cell line was performed using GAPDH mRNA transcription rates as endogenous control for normalization of expression levels. For Western blotting, P2X7R and Oct-4 expression levels were obtained and analyzed by densimetric analysis of protein bands and were compared to β-actin expression levels. The P2X7R was identified by two antibodies, which recognize the extracellular or C-terminus domain. Bars represent mean ± standard errors (S.E.) of three independent experiments performed in triplicate. Data were analyzed for statistical relevance with the One-Way ANOVA test followed by the Bonferroni post hoc test (*p<0,05,**p<0,01, ***p<0.001 compared to control data).

    Journal: PLoS ONE

    Article Title: Modulation of Mouse Embryonic Stem Cell Proliferation and Neural Differentiation by the P2X7 Receptor

    doi: 10.1371/journal.pone.0096281

    Figure Lengend Snippet: P2X7 receptor and Oct-4 expression levels in undifferentiated (und) and cells induced to differentiation (days 0–8) were determined by ( A ) real-time PCR and ( B,C ) Western blotting assays as described in Materials and . For real-time PCR, quantitative analysis of the relative expression of P2X7R and Oct-4 in E14Tg2A cell line was performed using GAPDH mRNA transcription rates as endogenous control for normalization of expression levels. For Western blotting, P2X7R and Oct-4 expression levels were obtained and analyzed by densimetric analysis of protein bands and were compared to β-actin expression levels. The P2X7R was identified by two antibodies, which recognize the extracellular or C-terminus domain. Bars represent mean ± standard errors (S.E.) of three independent experiments performed in triplicate. Data were analyzed for statistical relevance with the One-Way ANOVA test followed by the Bonferroni post hoc test (*p<0,05,**p<0,01, ***p<0.001 compared to control data).

    Article Snippet: The membranes were then incubated with primary antibodies for Oct-4 (polyclonal rabbit 1∶1000 Millipore), P2X7 receptor extracellular epitope (monoclonal rabbit 1∶2000 AbCam), P2X7 receptor C-terminus epitope (polyclonal rabbit 1∶1000 Alomone) and α-actin (1∶1000 Sigma-Aldrich) overnight at 4°C.

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

    ( A ) P2X7 receptor isoform A, B, C and k expression in undifferentiated (UND) and cells following 8 days of neural differentiation (DIFF) was determined by RT-PCR. ( B ) P2X7R isoforms expression levels were obtained and analyzed by densimetric analysis of DNA bands and were compared to GAPDH expression levels. Data were analyzed for statistical relevance with the One-Way ANOVA test followed by the Bonferroni post hoc test (*p<0,05,**p<0,01, ***p<0.001 compared to control data).

    Journal: PLoS ONE

    Article Title: Modulation of Mouse Embryonic Stem Cell Proliferation and Neural Differentiation by the P2X7 Receptor

    doi: 10.1371/journal.pone.0096281

    Figure Lengend Snippet: ( A ) P2X7 receptor isoform A, B, C and k expression in undifferentiated (UND) and cells following 8 days of neural differentiation (DIFF) was determined by RT-PCR. ( B ) P2X7R isoforms expression levels were obtained and analyzed by densimetric analysis of DNA bands and were compared to GAPDH expression levels. Data were analyzed for statistical relevance with the One-Way ANOVA test followed by the Bonferroni post hoc test (*p<0,05,**p<0,01, ***p<0.001 compared to control data).

    Article Snippet: The membranes were then incubated with primary antibodies for Oct-4 (polyclonal rabbit 1∶1000 Millipore), P2X7 receptor extracellular epitope (monoclonal rabbit 1∶2000 AbCam), P2X7 receptor C-terminus epitope (polyclonal rabbit 1∶1000 Alomone) and α-actin (1∶1000 Sigma-Aldrich) overnight at 4°C.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

    (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