aqp4  (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
    Name:
    Anti Aquaporin 4 AQP4 300 314 Antibody
    Description:
    Anti Aquaporin 4 AQP4 300 314 Antibody AQP 014 is a highly specific antibody directed against an epitope of rat AQP4 The antibody can be used in western blot immunohistochemistry and immunocytochemistry applications It has been designed to recognize the Aquaporin 4 channel from rat mouse and human samples
    Catalog Number:
    AQP-014
    Price:
    495.0
    Category:
    Primary Antibody
    Applications:
    Immunocytochemistry, Immunofluorescence, Immunohistochemistry, Western Blot
    Purity:
    Affinity purified on immobilized antigen.
    Immunogen:
    Synthetic peptide
    Size:
    50 mcl
    Antibody Type:
    Polyclonal Primary Antibodies
    Format:
    Lyophilized Powder
    Host:
    Rabbit
    Isotype:
    Rabbit IgG
    Buy from Supplier


    Structured Review

    Alomone Labs aqp4
    Anti Aquaporin 4 AQP4 300 314 Antibody
    Anti Aquaporin 4 AQP4 300 314 Antibody AQP 014 is a highly specific antibody directed against an epitope of rat AQP4 The antibody can be used in western blot immunohistochemistry and immunocytochemistry applications It has been designed to recognize the Aquaporin 4 channel from rat mouse and human samples
    https://www.bioz.com/result/aqp4/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    aqp4 - by Bioz Stars, 2021-09
    94/100 stars

    Images

    1) Product Images from "Glutamate and γ-aminobutyric acid differentially modulate glymphatic clearance of amyloid β through pulsation- and aquaporin-4 dependent mechanisms"

    Article Title: Glutamate and γ-aminobutyric acid differentially modulate glymphatic clearance of amyloid β through pulsation- and aquaporin-4 dependent mechanisms

    Journal: bioRxiv

    doi: 10.1101/2020.01.31.928481

    Hypertension induces brain Aβ deposition and glymphatic clearance impairment. (a–d) In vivo imaging of Aβ deposition in the cerebral cortex. Aβ deposition (FSB, green) was distinct along the vessels. (e) Enlarged VRS were observed. (f) 3D reconstruction of the vasculature in hypertension mice. The warping vessel is magnified on the right panel. (g-j) Ang-II evoked a significant increase in systolic blood pressure (SBP) and diastolic blood pressure (DBP) with no changes in heart rates (HR) and body weight (k-m). Glymphatic clearance impairment was evident in hypertension models. (n-o) Arterial diameters remained unchanged in hypertension models while vascular pulsatility was severely reduced (n=8-9 vessels per group) (s) . (q) Representative images of GFAP expression and AQP4 polarization in the cortex. No distinctive changes in AQP4 polarization (t) and GFAP expression (u) were observed in the hypertension model. (p, upper panel) Representative images of smooth muscle actin (SMA) and collagen expression in the cortex. No significant changes in SMA expression (r) and greater deposition of collagen (r) in vascular walls were observed in hypertension models. (p, lower panel) Immunology of Aβ 1–40 and Aβ 1–42 in hypertension model mice. Significant deposition of Aβ 1–40, but not Aβ 1–42, in vessels was observed. (p, lower panel) Co-labeling of collagen and Aβ 1–40 in hypertension. Aβ 1–40 co-localized with collagen (n=5-6 mice per group).
    Figure Legend Snippet: Hypertension induces brain Aβ deposition and glymphatic clearance impairment. (a–d) In vivo imaging of Aβ deposition in the cerebral cortex. Aβ deposition (FSB, green) was distinct along the vessels. (e) Enlarged VRS were observed. (f) 3D reconstruction of the vasculature in hypertension mice. The warping vessel is magnified on the right panel. (g-j) Ang-II evoked a significant increase in systolic blood pressure (SBP) and diastolic blood pressure (DBP) with no changes in heart rates (HR) and body weight (k-m). Glymphatic clearance impairment was evident in hypertension models. (n-o) Arterial diameters remained unchanged in hypertension models while vascular pulsatility was severely reduced (n=8-9 vessels per group) (s) . (q) Representative images of GFAP expression and AQP4 polarization in the cortex. No distinctive changes in AQP4 polarization (t) and GFAP expression (u) were observed in the hypertension model. (p, upper panel) Representative images of smooth muscle actin (SMA) and collagen expression in the cortex. No significant changes in SMA expression (r) and greater deposition of collagen (r) in vascular walls were observed in hypertension models. (p, lower panel) Immunology of Aβ 1–40 and Aβ 1–42 in hypertension model mice. Significant deposition of Aβ 1–40, but not Aβ 1–42, in vessels was observed. (p, lower panel) Co-labeling of collagen and Aβ 1–40 in hypertension. Aβ 1–40 co-localized with collagen (n=5-6 mice per group).

    Techniques Used: In Vivo Imaging, Mouse Assay, Expressing, Labeling

    Impairment of glymphatic clearance and deposition of Aβ plaques in APP-PS1 mice. (a) In vivo imaging of Aβ deposition in the cerebral cortex (FSB: green). Aβ deposition was evident in the parenchyma with no distinct CAA. (b) Immunology of Aβ 1–40 and Aβ 1–42 in APP-PS1. Significant numbers of Aβ 1–42-labeled amyloid plaques were observed in the parenchyma, but no marked deposition of Aβ 1–40. (c) Representative images of paravascular CSF tracer clearance at 100 μm below the cortical surface in APP-PS1 indicating severe impairment in penetration of fluorescence tracer (e) while no changes in paravascular movement was observed (d). (h-i) Expression of AQP4 and GFAP in cortex and hippocampus. Compared with WT control mice, APP-PS1 mice displayed significant decrease in AQP4 polarization and exhibited a marked increase in GFAP expression in the cortex (n=6 mice per group). No significant pulsatility changes were observed between APP-PS1 and WT control mice (f) (n=7-8 vessels per group).
    Figure Legend Snippet: Impairment of glymphatic clearance and deposition of Aβ plaques in APP-PS1 mice. (a) In vivo imaging of Aβ deposition in the cerebral cortex (FSB: green). Aβ deposition was evident in the parenchyma with no distinct CAA. (b) Immunology of Aβ 1–40 and Aβ 1–42 in APP-PS1. Significant numbers of Aβ 1–42-labeled amyloid plaques were observed in the parenchyma, but no marked deposition of Aβ 1–40. (c) Representative images of paravascular CSF tracer clearance at 100 μm below the cortical surface in APP-PS1 indicating severe impairment in penetration of fluorescence tracer (e) while no changes in paravascular movement was observed (d). (h-i) Expression of AQP4 and GFAP in cortex and hippocampus. Compared with WT control mice, APP-PS1 mice displayed significant decrease in AQP4 polarization and exhibited a marked increase in GFAP expression in the cortex (n=6 mice per group). No significant pulsatility changes were observed between APP-PS1 and WT control mice (f) (n=7-8 vessels per group).

    Techniques Used: Mouse Assay, In Vivo Imaging, Labeling, Fluorescence, Expressing

    Effects of Glutamate and GABA on tracer penetration into parenchyma are AQP4-dependent. (a) In vivo two-photon imaging of tracer clearance through the glymphatic pathway after intracisternal injection of GABA/glutamate and the respective antagonists in AQP4 -/- mice. (b, c) Quantification of CSF tracer influx into the surrounding parenchyma via 3D reconstruction and paravascular CSF tracer clearance at 100 μm below the cortical surface (d, e) . APV strongly accelerated paravasular movement while no significant changes were observed in mice receiving glutamate and CNQX. GABA and bicuculline did not affect paravascular movement. No significant changes in tracer penetration into the interstitium were observed among the experimental groups, compared with the vehicle group (n=6 mice per group).
    Figure Legend Snippet: Effects of Glutamate and GABA on tracer penetration into parenchyma are AQP4-dependent. (a) In vivo two-photon imaging of tracer clearance through the glymphatic pathway after intracisternal injection of GABA/glutamate and the respective antagonists in AQP4 -/- mice. (b, c) Quantification of CSF tracer influx into the surrounding parenchyma via 3D reconstruction and paravascular CSF tracer clearance at 100 μm below the cortical surface (d, e) . APV strongly accelerated paravasular movement while no significant changes were observed in mice receiving glutamate and CNQX. GABA and bicuculline did not affect paravascular movement. No significant changes in tracer penetration into the interstitium were observed among the experimental groups, compared with the vehicle group (n=6 mice per group).

    Techniques Used: In Vivo, Imaging, Injection, Mouse Assay

    Related Articles

    Labeling:

    Article Title: Colony-stimulating factor 1 receptor inhibition prevents disruption of the blood-retina barrier during chronic inflammation
    Article Snippet: .. For the labeling of neutrophils and aquaporin 4 (AQP4), slides were treated as above, and after the antigen retrieval, they were blocked for 30 min with 1x PBS, 5% NGS, 0.1% Triton X-100 and incubated overnight at 4 °C with a rabbit monoclonal antibody against myeloperoxidase (MPO, 1:1000, ab208670, Abcam, Cambridge, UK) or a rabbit polyclonal antibody against AQP4 (1:500, AQP-014, Alomone labs, Jerusalem, Israel). ..

    Next-Generation Sequencing:

    Article Title: Colony-stimulating factor 1 receptor inhibition prevents disruption of the blood-retina barrier during chronic inflammation
    Article Snippet: .. For the labeling of neutrophils and aquaporin 4 (AQP4), slides were treated as above, and after the antigen retrieval, they were blocked for 30 min with 1x PBS, 5% NGS, 0.1% Triton X-100 and incubated overnight at 4 °C with a rabbit monoclonal antibody against myeloperoxidase (MPO, 1:1000, ab208670, Abcam, Cambridge, UK) or a rabbit polyclonal antibody against AQP4 (1:500, AQP-014, Alomone labs, Jerusalem, Israel). ..

    Incubation:

    Article Title: Colony-stimulating factor 1 receptor inhibition prevents disruption of the blood-retina barrier during chronic inflammation
    Article Snippet: .. For the labeling of neutrophils and aquaporin 4 (AQP4), slides were treated as above, and after the antigen retrieval, they were blocked for 30 min with 1x PBS, 5% NGS, 0.1% Triton X-100 and incubated overnight at 4 °C with a rabbit monoclonal antibody against myeloperoxidase (MPO, 1:1000, ab208670, Abcam, Cambridge, UK) or a rabbit polyclonal antibody against AQP4 (1:500, AQP-014, Alomone labs, Jerusalem, Israel). ..

    Article Title: An in vitro bioengineered model of the human arterial neurovascular unit to study neurodegenerative diseases
    Article Snippet: .. For immunofluorescence, cryopreserved sections (arterial NVU and human brain sections) and fresh arterial NVU were blocked in 5% donkey serum and 1% BSA in PBS for 30 min at RT, incubated overnight at 4 °C with specific antibodies against the endothelial markers platelet endothelial cell adhesion molecule (CD31, RRID: AB_31432, WM59 Biolegend, 1:50) and von Willebrand factor (vWF, RRID:AB_259543, SigmaAldrich, 1:200), the SMC marker α-SM-actin (αSMA RRID:AB_476856, 1A4 SigmaAldrich, 1:200), the astrocyte markers glial fibrillary acidic protein (GFAP, RRID: AB_880203, Abcam, 1:200), and aquaporin 4 (AQ4, RRDI:AB_2039734, Alomone Labs, 1:100), the neuronal markers microtubule-associated protein 2 (MAP 2, RRID:AB_776174, Abcam, 1:200), β-tubulin III ( β-tub III, RRID:AB_2256751, Tuj1, 1:200), synapsin I (Syn, RRID:AB_2200097, Abcam, 1:200), Aβ markers Aβ 1–16 (6E10, RRID: AB_2565328, ThermoFisher Scientific, 1:50) and Aβ fibrils (OC fibril, RRID: AB_1977024, EMD Millipore, AB2286, 1:200) and phospho-Tau (AT8, RRID:AB_223648, ThermoFisher, 1:250). ..

    Article Title: A Synthesized Glucocorticoid- Induced Leucine Zipper Peptide Inhibits Retinal Müller Cell Gliosis
    Article Snippet: .. The membranes were blocked in 5% non-fat milk at room temperature for 1 h and incubated with the following antibodies: anti-monocyte chemoattractant protein (MCP)-1 (ab25124; Abcam, Cambridge, United Kingdom), anti-intercellular adhesion molecule (ICAM)-1 (ab171123; Proteintech, Chicago, IL, United States), anti-IL1β (ab9787; Abcam), Anti-tumor necrosis factor (TNF)α (PB0270, Boster Biological Technology, Wuhan, China), rabbit anti-p65 polyclonal antibody (ab16502; Abcam), anti-phospho-NF-κB p65 (Ser536) rabbit monoclonal antibody (3033P; Cell Signaling Technology, Beverly, MA, United States), anti-AQP4 (300-314, Alomone Labs, Jerusalem, Israel), anti-glial fibrillary acidic protein (GFAP) (ab10062, Abcam), or rabbit anti β-actin antibody (ab69512; Abcam) overnight. ..

    Article Title: Differential roles of VPS and RAAS in water homeostasis and a risk for kidney dysfunction in rats undergoing rapid fasting/dehydration with regular exercise
    Article Snippet: .. After blocking with a blocking solution (AQPs, 5% skimmed milk; caspase‐3, Blocking One [Nacalai Tesque]), the membranes were incubated overnight with the rabbit primary antibody anti‐AQP2 (1:5,000 dilution) (Alomone Labs, Jerusalem, Israel), anti‐AQP3 (1:10,000 dilution) (Alomone Labs), anti‐AQP4 (1:10,000 dilution) (Alomone Labs), or anti‐caspase‐3 (1:2000) (Cell Signaling Technology, Beverly, MA, USA) at 4°C with gentle shaking. ..

    Immunofluorescence:

    Article Title: An in vitro bioengineered model of the human arterial neurovascular unit to study neurodegenerative diseases
    Article Snippet: .. For immunofluorescence, cryopreserved sections (arterial NVU and human brain sections) and fresh arterial NVU were blocked in 5% donkey serum and 1% BSA in PBS for 30 min at RT, incubated overnight at 4 °C with specific antibodies against the endothelial markers platelet endothelial cell adhesion molecule (CD31, RRID: AB_31432, WM59 Biolegend, 1:50) and von Willebrand factor (vWF, RRID:AB_259543, SigmaAldrich, 1:200), the SMC marker α-SM-actin (αSMA RRID:AB_476856, 1A4 SigmaAldrich, 1:200), the astrocyte markers glial fibrillary acidic protein (GFAP, RRID: AB_880203, Abcam, 1:200), and aquaporin 4 (AQ4, RRDI:AB_2039734, Alomone Labs, 1:100), the neuronal markers microtubule-associated protein 2 (MAP 2, RRID:AB_776174, Abcam, 1:200), β-tubulin III ( β-tub III, RRID:AB_2256751, Tuj1, 1:200), synapsin I (Syn, RRID:AB_2200097, Abcam, 1:200), Aβ markers Aβ 1–16 (6E10, RRID: AB_2565328, ThermoFisher Scientific, 1:50) and Aβ fibrils (OC fibril, RRID: AB_1977024, EMD Millipore, AB2286, 1:200) and phospho-Tau (AT8, RRID:AB_223648, ThermoFisher, 1:250). ..

    Marker:

    Article Title: An in vitro bioengineered model of the human arterial neurovascular unit to study neurodegenerative diseases
    Article Snippet: .. For immunofluorescence, cryopreserved sections (arterial NVU and human brain sections) and fresh arterial NVU were blocked in 5% donkey serum and 1% BSA in PBS for 30 min at RT, incubated overnight at 4 °C with specific antibodies against the endothelial markers platelet endothelial cell adhesion molecule (CD31, RRID: AB_31432, WM59 Biolegend, 1:50) and von Willebrand factor (vWF, RRID:AB_259543, SigmaAldrich, 1:200), the SMC marker α-SM-actin (αSMA RRID:AB_476856, 1A4 SigmaAldrich, 1:200), the astrocyte markers glial fibrillary acidic protein (GFAP, RRID: AB_880203, Abcam, 1:200), and aquaporin 4 (AQ4, RRDI:AB_2039734, Alomone Labs, 1:100), the neuronal markers microtubule-associated protein 2 (MAP 2, RRID:AB_776174, Abcam, 1:200), β-tubulin III ( β-tub III, RRID:AB_2256751, Tuj1, 1:200), synapsin I (Syn, RRID:AB_2200097, Abcam, 1:200), Aβ markers Aβ 1–16 (6E10, RRID: AB_2565328, ThermoFisher Scientific, 1:50) and Aβ fibrils (OC fibril, RRID: AB_1977024, EMD Millipore, AB2286, 1:200) and phospho-Tau (AT8, RRID:AB_223648, ThermoFisher, 1:250). ..

    Injection:

    Article Title: Type I interferon-activated microglia are critical for neuromyelitis optica pathology.
    Article Snippet: .. Neuromyelitis optica (NMO) is an inflammatory disease of the central nervous system (CNS) most frequently mediated by serum autoantibodies against the water channel aquaporin 4, expressed on CNS astrocytes, resulting in primary astrocytopathy.. There is no cure for NMO, and treatment with Type I interferon (IFNI)-IFNβ is ineffective or even detrimental. ..

    Purification:

    Article Title: Type I interferon-activated microglia are critical for neuromyelitis optica pathology.
    Article Snippet: .. Neuromyelitis optica (NMO) is an inflammatory disease of the central nervous system (CNS) most frequently mediated by serum autoantibodies against the water channel aquaporin 4, expressed on CNS astrocytes, resulting in primary astrocytopathy.. There is no cure for NMO, and treatment with Type I interferon (IFNI)-IFNβ is ineffective or even detrimental. ..

    Blocking Assay:

    Article Title: Differential roles of VPS and RAAS in water homeostasis and a risk for kidney dysfunction in rats undergoing rapid fasting/dehydration with regular exercise
    Article Snippet: .. After blocking with a blocking solution (AQPs, 5% skimmed milk; caspase‐3, Blocking One [Nacalai Tesque]), the membranes were incubated overnight with the rabbit primary antibody anti‐AQP2 (1:5,000 dilution) (Alomone Labs, Jerusalem, Israel), anti‐AQP3 (1:10,000 dilution) (Alomone Labs), anti‐AQP4 (1:10,000 dilution) (Alomone Labs), or anti‐caspase‐3 (1:2000) (Cell Signaling Technology, Beverly, MA, USA) at 4°C with gentle shaking. ..

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    Alomone Labs anti asic1
    <t>ASIC1</t> and ASIC2a protein expression during OGD-Rep in three types of neurons. Cells were treated with OGD-Rep and prepared for immunofluorescence and western blot analysis. A, C. Representative pictures and summary data showing expressions of ASIC1 and ASIC2a using immunofluorescence. B, D. Representative bands and summary data of ASIC1 and ASIC2a expression in western blotting. Data are the mean ± SD in each group. *P
    Anti Asic1, 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 asic1/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti asic1 - by Bioz Stars, 2021-09
    94/100 stars
      Buy from Supplier

    94
    Alomone Labs anti aqp4
    The synthesized GILZ-p inhibited LPS induced Müller cell gliosis. Western blot analysis was performed to determine the protein expression levels of glial fibrillary acidic protein (GFAP) (A,B) , and <t>AQP4</t> (C,D) in Müller cells treated with 1000 ng/ml LPS in combination with different concentrations of GILZ-p (0.01, 0.1, 1, and 10 μM) for 24 h. β-actin was used as the loading control. The results of quantitative analysis, as determined by densitometric analysis, were expressed as relative to β-actin. Data represent the mean ± SE; the Mann–Whitney U -test was used for comparisons between two groups. n = 3 for each group. ∗ P
    Anti Aqp4, 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 aqp4/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti aqp4 - by Bioz Stars, 2021-09
    94/100 stars
      Buy from Supplier

    94
    Alomone Labs anti cxcr4
    Effects of AMD3100 on prazosin and cyclazosin induced ERK1/2 phosphorylation. ERK1/2 phosphorylation was measured and analyzed as in Fig 3 . Cells were pre- incubated with AMD3100 (10 μM, 15 min), followed by stimulation with vehicle (control, ctrl), prazosin (Praz) or cyclazosin (Cycl, 100 μM each) for 20 min. A. Representative images from Western blot experiments with cells transfected with <t>CXCR4.</t> The migration position of molecular mass standards is indicated. B. Densitometric quantification of the band intensities, expressed as pERK1/2/ERK1/2, from n = 5 independent experiments as in A. Data are mean ± SE. *: p
    Anti Cxcr4, 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 cxcr4/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cxcr4 - by Bioz Stars, 2021-09
    94/100 stars
      Buy from Supplier

    Image Search Results


    ASIC1 and ASIC2a protein expression during OGD-Rep in three types of neurons. Cells were treated with OGD-Rep and prepared for immunofluorescence and western blot analysis. A, C. Representative pictures and summary data showing expressions of ASIC1 and ASIC2a using immunofluorescence. B, D. Representative bands and summary data of ASIC1 and ASIC2a expression in western blotting. Data are the mean ± SD in each group. *P

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Down-regulation of ASICs current and the calcium transients by disrupting PICK1 protects primary cultured mouse cortical neurons from OGD-Rep insults

    doi:

    Figure Lengend Snippet: ASIC1 and ASIC2a protein expression during OGD-Rep in three types of neurons. Cells were treated with OGD-Rep and prepared for immunofluorescence and western blot analysis. A, C. Representative pictures and summary data showing expressions of ASIC1 and ASIC2a using immunofluorescence. B, D. Representative bands and summary data of ASIC1 and ASIC2a expression in western blotting. Data are the mean ± SD in each group. *P

    Article Snippet: Cerebral cortical neurons were collected and fixed with 4% paraformaldehyde in PBS for 30 min. Then permeabilized with PBS/0.3% Triton X-100 for 30 min, and all cells were blocked with 2% goat serum and 1% bovine serum albumin (BSA) in PBS for 1 h, and then incubated with anti-ASIC1 (Alomone labs, Israel) or anti-ASIC2a (Alomone labs, Israel) overnight at 4°C.

    Techniques: Expressing, Immunofluorescence, Western Blot

    The synthesized GILZ-p inhibited LPS induced Müller cell gliosis. Western blot analysis was performed to determine the protein expression levels of glial fibrillary acidic protein (GFAP) (A,B) , and AQP4 (C,D) in Müller cells treated with 1000 ng/ml LPS in combination with different concentrations of GILZ-p (0.01, 0.1, 1, and 10 μM) for 24 h. β-actin was used as the loading control. The results of quantitative analysis, as determined by densitometric analysis, were expressed as relative to β-actin. Data represent the mean ± SE; the Mann–Whitney U -test was used for comparisons between two groups. n = 3 for each group. ∗ P

    Journal: Frontiers in Pharmacology

    Article Title: A Synthesized Glucocorticoid- Induced Leucine Zipper Peptide Inhibits Retinal Müller Cell Gliosis

    doi: 10.3389/fphar.2018.00331

    Figure Lengend Snippet: The synthesized GILZ-p inhibited LPS induced Müller cell gliosis. Western blot analysis was performed to determine the protein expression levels of glial fibrillary acidic protein (GFAP) (A,B) , and AQP4 (C,D) in Müller cells treated with 1000 ng/ml LPS in combination with different concentrations of GILZ-p (0.01, 0.1, 1, and 10 μM) for 24 h. β-actin was used as the loading control. The results of quantitative analysis, as determined by densitometric analysis, were expressed as relative to β-actin. Data represent the mean ± SE; the Mann–Whitney U -test was used for comparisons between two groups. n = 3 for each group. ∗ P

    Article Snippet: The membranes were blocked in 5% non-fat milk at room temperature for 1 h and incubated with the following antibodies: anti-monocyte chemoattractant protein (MCP)-1 (ab25124; Abcam, Cambridge, United Kingdom), anti-intercellular adhesion molecule (ICAM)-1 (ab171123; Proteintech, Chicago, IL, United States), anti-IL1β (ab9787; Abcam), Anti-tumor necrosis factor (TNF)α (PB0270, Boster Biological Technology, Wuhan, China), rabbit anti-p65 polyclonal antibody (ab16502; Abcam), anti-phospho-NF-κB p65 (Ser536) rabbit monoclonal antibody (3033P; Cell Signaling Technology, Beverly, MA, United States), anti-AQP4 (300-314, Alomone Labs, Jerusalem, Israel), anti-glial fibrillary acidic protein (GFAP) (ab10062, Abcam), or rabbit anti β-actin antibody (ab69512; Abcam) overnight.

    Techniques: Synthesized, Western Blot, Expressing, MANN-WHITNEY

    Alteration of GFAP and AQP4 in the cortex of EAE mice. A , Alterations of astrocytic proteins in the cortex at different stages of rrEAE. B , Summary of the alterations of intensities. C , Summary of increased GFAP + cells. (B, C) One-Way ANOVA followed by Fisher’s test. *, significant difference from Control for each antibody, p

    Journal: Acta Neuropathologica Communications

    Article Title: Astrocytes differentially respond to inflammatory autoimmune insults and imbalances of neural activity

    doi: 10.1186/2051-5960-1-70

    Figure Lengend Snippet: Alteration of GFAP and AQP4 in the cortex of EAE mice. A , Alterations of astrocytic proteins in the cortex at different stages of rrEAE. B , Summary of the alterations of intensities. C , Summary of increased GFAP + cells. (B, C) One-Way ANOVA followed by Fisher’s test. *, significant difference from Control for each antibody, p

    Article Snippet: The following antibodies were used in our study: rabbit polyclonal anti-Kv1.4, anti-AQP4 (Alomone Labs, Jerusalem, Israel), and anti-PKCγ (Santa Cruz Biotechnology, Dallas, TX); goat polyclonal anti-GFAP (AbCAM, Cambridge, MA, USA); chicken polyclonal anti-Vimentin (Millipore, Temecula, CA); and Dylight 488-, Dylight 649-, Cy3-, and Cy5-conjugated secondary antibodies (Jackson Immuno Research Laboratories, West Grove, PA, USA).

    Techniques: Mouse Assay

    Altered astrocytes in the hippocampus and cortex in Kv3.1 KO mice. The confocal image stacks of hippocampus (A-C) and cortex (D-F) were costained for GFAP (green) and AQP4 (red) from WT (A,D) , AnkG KO (B,E) and Kv3.1 KO (C,F) mice. The collapsed 2D image is on the top and 3 cross sections are at the bottom. The crossbars are centered on astrocyte endfeet with colocalizing AQP4 and GFAP. Scale bars, 100 μm.

    Journal: Acta Neuropathologica Communications

    Article Title: Astrocytes differentially respond to inflammatory autoimmune insults and imbalances of neural activity

    doi: 10.1186/2051-5960-1-70

    Figure Lengend Snippet: Altered astrocytes in the hippocampus and cortex in Kv3.1 KO mice. The confocal image stacks of hippocampus (A-C) and cortex (D-F) were costained for GFAP (green) and AQP4 (red) from WT (A,D) , AnkG KO (B,E) and Kv3.1 KO (C,F) mice. The collapsed 2D image is on the top and 3 cross sections are at the bottom. The crossbars are centered on astrocyte endfeet with colocalizing AQP4 and GFAP. Scale bars, 100 μm.

    Article Snippet: The following antibodies were used in our study: rabbit polyclonal anti-Kv1.4, anti-AQP4 (Alomone Labs, Jerusalem, Israel), and anti-PKCγ (Santa Cruz Biotechnology, Dallas, TX); goat polyclonal anti-GFAP (AbCAM, Cambridge, MA, USA); chicken polyclonal anti-Vimentin (Millipore, Temecula, CA); and Dylight 488-, Dylight 649-, Cy3-, and Cy5-conjugated secondary antibodies (Jackson Immuno Research Laboratories, West Grove, PA, USA).

    Techniques: Mouse Assay

    Differentially altered expression of astrocytic proteins in the cerebellum of EAE mice. A , The confocal image stack of cerebellar molecular layer that was stained for GFAP (green), AQP4 (red) and Vim (blue) from a control mouse. Collapsed 2D image is on the left and 3 cross sections of 3D are on the right. B , The images at the peak stage of an rrEAE mouse. C , The images at the remitting stage of an rrEAE mouse. D , The images at the relapsing stage of an rrEAE mouse. E , The confocal images of cerebellar WM from a Thy1-YFP transgenic mouse. YFP (green), AQP4 (red) and GFAP (blue). In (A-E) , the crossbars are centered on astrocytic endfeet with colocalizing AQP4 and GFAP. F , The confocal images at the peak stage of chEAE. The crossbars show the lesion edge with upregulated AQP4 and GFAP. G , Structural diagram of cerebellar cortex. H , Summary of changes of protein levels during rrEAE in cerebellar molecular layer. One-Way ANOVA followed by Fisher’s test. *, significant difference from Control for each antibody, p

    Journal: Acta Neuropathologica Communications

    Article Title: Astrocytes differentially respond to inflammatory autoimmune insults and imbalances of neural activity

    doi: 10.1186/2051-5960-1-70

    Figure Lengend Snippet: Differentially altered expression of astrocytic proteins in the cerebellum of EAE mice. A , The confocal image stack of cerebellar molecular layer that was stained for GFAP (green), AQP4 (red) and Vim (blue) from a control mouse. Collapsed 2D image is on the left and 3 cross sections of 3D are on the right. B , The images at the peak stage of an rrEAE mouse. C , The images at the remitting stage of an rrEAE mouse. D , The images at the relapsing stage of an rrEAE mouse. E , The confocal images of cerebellar WM from a Thy1-YFP transgenic mouse. YFP (green), AQP4 (red) and GFAP (blue). In (A-E) , the crossbars are centered on astrocytic endfeet with colocalizing AQP4 and GFAP. F , The confocal images at the peak stage of chEAE. The crossbars show the lesion edge with upregulated AQP4 and GFAP. G , Structural diagram of cerebellar cortex. H , Summary of changes of protein levels during rrEAE in cerebellar molecular layer. One-Way ANOVA followed by Fisher’s test. *, significant difference from Control for each antibody, p

    Article Snippet: The following antibodies were used in our study: rabbit polyclonal anti-Kv1.4, anti-AQP4 (Alomone Labs, Jerusalem, Israel), and anti-PKCγ (Santa Cruz Biotechnology, Dallas, TX); goat polyclonal anti-GFAP (AbCAM, Cambridge, MA, USA); chicken polyclonal anti-Vimentin (Millipore, Temecula, CA); and Dylight 488-, Dylight 649-, Cy3-, and Cy5-conjugated secondary antibodies (Jackson Immuno Research Laboratories, West Grove, PA, USA).

    Techniques: Expressing, Mouse Assay, Staining, Transgenic Assay

    Activation of astrocytes in the hippocampus of EAE mice. A , Increased GFAP but not AQP4 staining in the hippocampus of EAE mice. B , Increased Vim staining in the hippocampus during EAE progression. C , Enlarged images of individual astrocytes in the hippocampus clearly show the increase of Vim staining. High magnification confocal image stacks were obtained from control (D) and EAE (E) Thy1-YFP transgenic mice. Images contain YFP (green), GFAP (blue) and AQP4 (red). The collapsed 2D image is on the left, and 3 cross sections are on the right. The crossbars reveal astrocytic endfeet with colocalizing AQP4 and GFAP. F , Summary of the levels of astrocytic proteins at different stages during EAE progression. One-Way ANOVA followed by Fisher’s test, *, significant difference from Control for each antibody, p

    Journal: Acta Neuropathologica Communications

    Article Title: Astrocytes differentially respond to inflammatory autoimmune insults and imbalances of neural activity

    doi: 10.1186/2051-5960-1-70

    Figure Lengend Snippet: Activation of astrocytes in the hippocampus of EAE mice. A , Increased GFAP but not AQP4 staining in the hippocampus of EAE mice. B , Increased Vim staining in the hippocampus during EAE progression. C , Enlarged images of individual astrocytes in the hippocampus clearly show the increase of Vim staining. High magnification confocal image stacks were obtained from control (D) and EAE (E) Thy1-YFP transgenic mice. Images contain YFP (green), GFAP (blue) and AQP4 (red). The collapsed 2D image is on the left, and 3 cross sections are on the right. The crossbars reveal astrocytic endfeet with colocalizing AQP4 and GFAP. F , Summary of the levels of astrocytic proteins at different stages during EAE progression. One-Way ANOVA followed by Fisher’s test, *, significant difference from Control for each antibody, p

    Article Snippet: The following antibodies were used in our study: rabbit polyclonal anti-Kv1.4, anti-AQP4 (Alomone Labs, Jerusalem, Israel), and anti-PKCγ (Santa Cruz Biotechnology, Dallas, TX); goat polyclonal anti-GFAP (AbCAM, Cambridge, MA, USA); chicken polyclonal anti-Vimentin (Millipore, Temecula, CA); and Dylight 488-, Dylight 649-, Cy3-, and Cy5-conjugated secondary antibodies (Jackson Immuno Research Laboratories, West Grove, PA, USA).

    Techniques: Activation Assay, Mouse Assay, Staining, Transgenic Assay

    Upregulation of GFAP and AQP4 in the cerebellum in AnkG and Kv3.1 KO mice. A , High magnification image stacks of cerebellar molecular layer stained with anti-PKCγ (green) and anti-GFAP (red) antibodies. The collapsed 2D image is on the left, and 3 cross sections are on the right. B , Confocal image stacks from the AnkG KO mice. C , Confocal image stacks from the Kv3.1 KO mice. In (A,C) , the crossbars reveal radially oriented GFAP + Bergmann glial processes in (A) WT and (C) Kv3.1 KO mice. In (B) , the crossbars are centered on highly upregulated GFAP + astrocyte processes in the absence of Purkinje neurons in an AnkG KO mouse. D , A single confocal image of the granule cell layer in a WT mouse. E , An image from the AnkG KO mice. F , An image from the Kv3.1 KO mice. G , Normalized fluorescence intensity in the molecular layer. H , Normalized fluorescence intensity in the granule cell layer. (G, H) One-Way ANOVA followed by Fisher’s test. *, significant difference from Wildtype for each antibody, p

    Journal: Acta Neuropathologica Communications

    Article Title: Astrocytes differentially respond to inflammatory autoimmune insults and imbalances of neural activity

    doi: 10.1186/2051-5960-1-70

    Figure Lengend Snippet: Upregulation of GFAP and AQP4 in the cerebellum in AnkG and Kv3.1 KO mice. A , High magnification image stacks of cerebellar molecular layer stained with anti-PKCγ (green) and anti-GFAP (red) antibodies. The collapsed 2D image is on the left, and 3 cross sections are on the right. B , Confocal image stacks from the AnkG KO mice. C , Confocal image stacks from the Kv3.1 KO mice. In (A,C) , the crossbars reveal radially oriented GFAP + Bergmann glial processes in (A) WT and (C) Kv3.1 KO mice. In (B) , the crossbars are centered on highly upregulated GFAP + astrocyte processes in the absence of Purkinje neurons in an AnkG KO mouse. D , A single confocal image of the granule cell layer in a WT mouse. E , An image from the AnkG KO mice. F , An image from the Kv3.1 KO mice. G , Normalized fluorescence intensity in the molecular layer. H , Normalized fluorescence intensity in the granule cell layer. (G, H) One-Way ANOVA followed by Fisher’s test. *, significant difference from Wildtype for each antibody, p

    Article Snippet: The following antibodies were used in our study: rabbit polyclonal anti-Kv1.4, anti-AQP4 (Alomone Labs, Jerusalem, Israel), and anti-PKCγ (Santa Cruz Biotechnology, Dallas, TX); goat polyclonal anti-GFAP (AbCAM, Cambridge, MA, USA); chicken polyclonal anti-Vimentin (Millipore, Temecula, CA); and Dylight 488-, Dylight 649-, Cy3-, and Cy5-conjugated secondary antibodies (Jackson Immuno Research Laboratories, West Grove, PA, USA).

    Techniques: Mouse Assay, Staining, Fluorescence

    Activation of astrocytes in spinal cord white matter. A , Clinical scores (top) and body weight (bottom) of mice with chEAE. B , White matter (WM) and gray matter (GM) in spinal cord longitudinal section was stained with FMG (green) and nuclear dye (blue). Box 1 shows both GM and WM and box 2 shows only WM. Spinal cord sections, control (C) and EAE peak (D) , were co-stained for GFAP (green, top), AQP4 (red), Hoechst (blue), and FMG (green, bottom). Co-staining of Kv1.4 (green, top), Vim (red), Hoechst (blue) and FMG (green, bottom) were also performed on control (E) and EAE (F) spinal cord sections. High magnification confocal image stacks were obtained from control (G) and EAE (H) Thy1-YFP transgenic mice. Images contain YFP (green), GFAP (blue) and AQP4 (red). The collapsed 2D image is on the left, and 3 cross sections are on the right. In (G) , the crossbars are centered on a putative node of Ranvier. In (H) , the crossbars are centered on the AQP4+/GFAP + lesion edge. Scale bars, 500 μm in C-F , 50 μm in G , H .

    Journal: Acta Neuropathologica Communications

    Article Title: Astrocytes differentially respond to inflammatory autoimmune insults and imbalances of neural activity

    doi: 10.1186/2051-5960-1-70

    Figure Lengend Snippet: Activation of astrocytes in spinal cord white matter. A , Clinical scores (top) and body weight (bottom) of mice with chEAE. B , White matter (WM) and gray matter (GM) in spinal cord longitudinal section was stained with FMG (green) and nuclear dye (blue). Box 1 shows both GM and WM and box 2 shows only WM. Spinal cord sections, control (C) and EAE peak (D) , were co-stained for GFAP (green, top), AQP4 (red), Hoechst (blue), and FMG (green, bottom). Co-staining of Kv1.4 (green, top), Vim (red), Hoechst (blue) and FMG (green, bottom) were also performed on control (E) and EAE (F) spinal cord sections. High magnification confocal image stacks were obtained from control (G) and EAE (H) Thy1-YFP transgenic mice. Images contain YFP (green), GFAP (blue) and AQP4 (red). The collapsed 2D image is on the left, and 3 cross sections are on the right. In (G) , the crossbars are centered on a putative node of Ranvier. In (H) , the crossbars are centered on the AQP4+/GFAP + lesion edge. Scale bars, 500 μm in C-F , 50 μm in G , H .

    Article Snippet: The following antibodies were used in our study: rabbit polyclonal anti-Kv1.4, anti-AQP4 (Alomone Labs, Jerusalem, Israel), and anti-PKCγ (Santa Cruz Biotechnology, Dallas, TX); goat polyclonal anti-GFAP (AbCAM, Cambridge, MA, USA); chicken polyclonal anti-Vimentin (Millipore, Temecula, CA); and Dylight 488-, Dylight 649-, Cy3-, and Cy5-conjugated secondary antibodies (Jackson Immuno Research Laboratories, West Grove, PA, USA).

    Techniques: Activation Assay, Mouse Assay, Staining, Transgenic Assay

    Effects of AMD3100 on prazosin and cyclazosin induced ERK1/2 phosphorylation. ERK1/2 phosphorylation was measured and analyzed as in Fig 3 . Cells were pre- incubated with AMD3100 (10 μM, 15 min), followed by stimulation with vehicle (control, ctrl), prazosin (Praz) or cyclazosin (Cycl, 100 μM each) for 20 min. A. Representative images from Western blot experiments with cells transfected with CXCR4. The migration position of molecular mass standards is indicated. B. Densitometric quantification of the band intensities, expressed as pERK1/2/ERK1/2, from n = 5 independent experiments as in A. Data are mean ± SE. *: p

    Journal: PLoS ONE

    Article Title: Partial agonist activity of α1-adrenergic receptor antagonists for chemokine (C-X-C motif) receptor 4 and atypical chemokine receptor 3

    doi: 10.1371/journal.pone.0204041

    Figure Lengend Snippet: Effects of AMD3100 on prazosin and cyclazosin induced ERK1/2 phosphorylation. ERK1/2 phosphorylation was measured and analyzed as in Fig 3 . Cells were pre- incubated with AMD3100 (10 μM, 15 min), followed by stimulation with vehicle (control, ctrl), prazosin (Praz) or cyclazosin (Cycl, 100 μM each) for 20 min. A. Representative images from Western blot experiments with cells transfected with CXCR4. The migration position of molecular mass standards is indicated. B. Densitometric quantification of the band intensities, expressed as pERK1/2/ERK1/2, from n = 5 independent experiments as in A. Data are mean ± SE. *: p

    Article Snippet: Cells were then blocked with 2% FBS in cold PBS for 30 min, followed by incubation with anti-CXCR4 (ACR-014, Alomone Labs) and anti-ACKR3 (MAB42273, R & D Systems) antibodies for 1h on ice.

    Techniques: Incubation, Western Blot, Transfection, Migration

    Screening of adrenergic receptor antagonists for CXCR4 and ACKR3 agonist activity in PRESTO-Tango β-arrestin recruitment assays. Data are mean ± SE from n = 4 independent experiments (in triplicates). Cells were stimulated with vehicle, 100 μM of individual AR antagonists or with 200 nM of CXCL12. Luminescence signals are expressed as fold of vehicle-treated cells (control, = 1). *: p

    Journal: PLoS ONE

    Article Title: Partial agonist activity of α1-adrenergic receptor antagonists for chemokine (C-X-C motif) receptor 4 and atypical chemokine receptor 3

    doi: 10.1371/journal.pone.0204041

    Figure Lengend Snippet: Screening of adrenergic receptor antagonists for CXCR4 and ACKR3 agonist activity in PRESTO-Tango β-arrestin recruitment assays. Data are mean ± SE from n = 4 independent experiments (in triplicates). Cells were stimulated with vehicle, 100 μM of individual AR antagonists or with 200 nM of CXCL12. Luminescence signals are expressed as fold of vehicle-treated cells (control, = 1). *: p

    Article Snippet: Cells were then blocked with 2% FBS in cold PBS for 30 min, followed by incubation with anti-CXCR4 (ACR-014, Alomone Labs) and anti-ACKR3 (MAB42273, R & D Systems) antibodies for 1h on ice.

    Techniques: Activity Assay

    Prazosin and cyclazosin induce ERK1/2 phosphorylation. HEK293 cells transfected with CXCR4 ( A/B ) or ACKR3 ( C/D ) were stimulated with CXCL12 (100 nM), prazosin (100 μM) or cyclazosin (100 μM) for various time periods as indicated. ERK1/2 phosphorylation was monitored by Western blotting of cell lysates with anti-phophoERK1/2 (pERK) and anti-ERK1/2 (ERK1/2). A. Representative images from Western blot experiments with cells transfected with CXCR4. The migration position of molecular mass standards is indicated. B. Densitometric quantification of the band intensities, expressed as pERK1/2/ERK1/2, from n = 3 independent experiments as in A. Data are mean ± SE. Symbols (*: CXCL12; #: cyclazosin; $: prazosin) indicate significant differences vs. t = 0 min. C. Representative images from Western blot experiments with cells transfected with ACKR3. The migration position of molecular mass standards is indicated. D. Densitometric quantification of the band intensities, expressed as pERK1/2/ERK1/2, from n = 5 independent experiments as in B. Data are mean ± SE. Symbols (*: CXCL12; #: cyclazosin; $: prazosin) indicate significant differences vs. t = 0 min.

    Journal: PLoS ONE

    Article Title: Partial agonist activity of α1-adrenergic receptor antagonists for chemokine (C-X-C motif) receptor 4 and atypical chemokine receptor 3

    doi: 10.1371/journal.pone.0204041

    Figure Lengend Snippet: Prazosin and cyclazosin induce ERK1/2 phosphorylation. HEK293 cells transfected with CXCR4 ( A/B ) or ACKR3 ( C/D ) were stimulated with CXCL12 (100 nM), prazosin (100 μM) or cyclazosin (100 μM) for various time periods as indicated. ERK1/2 phosphorylation was monitored by Western blotting of cell lysates with anti-phophoERK1/2 (pERK) and anti-ERK1/2 (ERK1/2). A. Representative images from Western blot experiments with cells transfected with CXCR4. The migration position of molecular mass standards is indicated. B. Densitometric quantification of the band intensities, expressed as pERK1/2/ERK1/2, from n = 3 independent experiments as in A. Data are mean ± SE. Symbols (*: CXCL12; #: cyclazosin; $: prazosin) indicate significant differences vs. t = 0 min. C. Representative images from Western blot experiments with cells transfected with ACKR3. The migration position of molecular mass standards is indicated. D. Densitometric quantification of the band intensities, expressed as pERK1/2/ERK1/2, from n = 5 independent experiments as in B. Data are mean ± SE. Symbols (*: CXCL12; #: cyclazosin; $: prazosin) indicate significant differences vs. t = 0 min.

    Article Snippet: Cells were then blocked with 2% FBS in cold PBS for 30 min, followed by incubation with anti-CXCR4 (ACR-014, Alomone Labs) and anti-ACKR3 (MAB42273, R & D Systems) antibodies for 1h on ice.

    Techniques: Transfection, Western Blot, Migration

    Prazosin and cyclazosin induce internalization of CXCR4 and ACKR3 in hVSMCs. A. hVSMC were treated with 100 μM cyclazosin or prazosin at 37°C for 15 or 30 min, stained with anti-CXCR4/Alexa 647-conjugated anti-rabbit and anti-ACKR3/Alexa 488-conjugated anti-mouse and analyzed for receptor expression via flow cytometry. RFU: relative fluorescence units. The horizontal and vertical lines show the gating thresholds for CXCR4 (Alexa 647) and ACKR3 (Alexa 488). B-E . Quantification of CXCR4 ( B/C ) and ACKR3 ( D/E ) cell surface expression by flow cytometry. Cells were stimulated with 100 μM cyclazosin ( B/D ) or prazosin ( C/E ) as indicated. Data are mean ± SE from 4 independent experiments. *: p

    Journal: PLoS ONE

    Article Title: Partial agonist activity of α1-adrenergic receptor antagonists for chemokine (C-X-C motif) receptor 4 and atypical chemokine receptor 3

    doi: 10.1371/journal.pone.0204041

    Figure Lengend Snippet: Prazosin and cyclazosin induce internalization of CXCR4 and ACKR3 in hVSMCs. A. hVSMC were treated with 100 μM cyclazosin or prazosin at 37°C for 15 or 30 min, stained with anti-CXCR4/Alexa 647-conjugated anti-rabbit and anti-ACKR3/Alexa 488-conjugated anti-mouse and analyzed for receptor expression via flow cytometry. RFU: relative fluorescence units. The horizontal and vertical lines show the gating thresholds for CXCR4 (Alexa 647) and ACKR3 (Alexa 488). B-E . Quantification of CXCR4 ( B/C ) and ACKR3 ( D/E ) cell surface expression by flow cytometry. Cells were stimulated with 100 μM cyclazosin ( B/D ) or prazosin ( C/E ) as indicated. Data are mean ± SE from 4 independent experiments. *: p

    Article Snippet: Cells were then blocked with 2% FBS in cold PBS for 30 min, followed by incubation with anti-CXCR4 (ACR-014, Alomone Labs) and anti-ACKR3 (MAB42273, R & D Systems) antibodies for 1h on ice.

    Techniques: Staining, Expressing, Flow Cytometry, Cytometry, Fluorescence

    Prazosin and cyclazosin induce chemical shift changes in the NMR spectra of CXCR4 and ACKR3 in membranes. 1 H- 13 C HSQC spectra of reductively methylated CXCR4 ( A-C ) and ACKR3 ( D-F ) membrane preparations were recorded without (black) and with (red) 200 μM prazosin ( A/D ), cyclazosin ( B/E ) or atipamezol ( C/F ). Black arrows indicate significant differences in chemical shifts or broadening (loss) of the signal.

    Journal: PLoS ONE

    Article Title: Partial agonist activity of α1-adrenergic receptor antagonists for chemokine (C-X-C motif) receptor 4 and atypical chemokine receptor 3

    doi: 10.1371/journal.pone.0204041

    Figure Lengend Snippet: Prazosin and cyclazosin induce chemical shift changes in the NMR spectra of CXCR4 and ACKR3 in membranes. 1 H- 13 C HSQC spectra of reductively methylated CXCR4 ( A-C ) and ACKR3 ( D-F ) membrane preparations were recorded without (black) and with (red) 200 μM prazosin ( A/D ), cyclazosin ( B/E ) or atipamezol ( C/F ). Black arrows indicate significant differences in chemical shifts or broadening (loss) of the signal.

    Article Snippet: Cells were then blocked with 2% FBS in cold PBS for 30 min, followed by incubation with anti-CXCR4 (ACR-014, Alomone Labs) and anti-ACKR3 (MAB42273, R & D Systems) antibodies for 1h on ice.

    Techniques: Nuclear Magnetic Resonance, Methylation

    CXCL12-inudced ERK1/2 phosphorylation in HEK293 cells is augmented after transfection with CXCR4 and ACKR3. HEK293 cells were transfected with empty vector (left), CXCR4 (center) or ACKR3 (right) and stimulated with 100 nM of CXCL12 for various time periods as indicated. ERK1/2 phosphorylation was monitored by Western blotting of cell lysates with anti-phophoERK1/2 (pERK) and anti-ERK1/2. The migration position of molecular mass standards is indicated.

    Journal: PLoS ONE

    Article Title: Partial agonist activity of α1-adrenergic receptor antagonists for chemokine (C-X-C motif) receptor 4 and atypical chemokine receptor 3

    doi: 10.1371/journal.pone.0204041

    Figure Lengend Snippet: CXCL12-inudced ERK1/2 phosphorylation in HEK293 cells is augmented after transfection with CXCR4 and ACKR3. HEK293 cells were transfected with empty vector (left), CXCR4 (center) or ACKR3 (right) and stimulated with 100 nM of CXCL12 for various time periods as indicated. ERK1/2 phosphorylation was monitored by Western blotting of cell lysates with anti-phophoERK1/2 (pERK) and anti-ERK1/2. The migration position of molecular mass standards is indicated.

    Article Snippet: Cells were then blocked with 2% FBS in cold PBS for 30 min, followed by incubation with anti-CXCR4 (ACR-014, Alomone Labs) and anti-ACKR3 (MAB42273, R & D Systems) antibodies for 1h on ice.

    Techniques: Transfection, Plasmid Preparation, Western Blot, Migration

    Dose-dependent effects of prazosin and cyclazosin in PRESTO-Tango β-arrestin recruitment assays for CXCR4 and ACKR3. Data are mean ± SE from n = 3 independent experiments (in triplicates). RLU: relative luminescence units. A-C: CXCR4 PRESTO-Tango assays. A. Grey circles: cells treated with cyclazosin. Black circles: cells treated with prazosin. B. Cells were pre-incubated with vehicle (control, grey circles) or AMD3100 (10 μM; black circles) for 15 min, followed by stimulation with cyclazosin. **: p

    Journal: PLoS ONE

    Article Title: Partial agonist activity of α1-adrenergic receptor antagonists for chemokine (C-X-C motif) receptor 4 and atypical chemokine receptor 3

    doi: 10.1371/journal.pone.0204041

    Figure Lengend Snippet: Dose-dependent effects of prazosin and cyclazosin in PRESTO-Tango β-arrestin recruitment assays for CXCR4 and ACKR3. Data are mean ± SE from n = 3 independent experiments (in triplicates). RLU: relative luminescence units. A-C: CXCR4 PRESTO-Tango assays. A. Grey circles: cells treated with cyclazosin. Black circles: cells treated with prazosin. B. Cells were pre-incubated with vehicle (control, grey circles) or AMD3100 (10 μM; black circles) for 15 min, followed by stimulation with cyclazosin. **: p

    Article Snippet: Cells were then blocked with 2% FBS in cold PBS for 30 min, followed by incubation with anti-CXCR4 (ACR-014, Alomone Labs) and anti-ACKR3 (MAB42273, R & D Systems) antibodies for 1h on ice.

    Techniques: Incubation