anti p2y receptor antibodies  (Alomone Labs)


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    Alomone Labs anti p2y receptor antibodies
    Expression of <t>P2Y</t> receptors in osteoblasts. Using flow cytometry analysis, P2Y 1 , P2Y 2 , P2Y 4 , P2Y 6 , P2Y 11 , P2Y 12 , and P2Y 13 receptors were detected in osteoblasts using their respective Alomone Labs antibodies ( open histograms ). Incubating the antibody with the control peptide antigen was used as a negative control ( filled histograms ). Adapted from reference 41 with permission of Elsevier
    Anti P2y Receptor Antibodies, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p2y receptor antibodies/product/Alomone Labs
    Average 91 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    anti p2y receptor antibodies - by Bioz Stars, 2022-12
    91/100 stars

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    1) Product Images from "Using antibodies against P2Y and P2X receptors in purinergic signaling research"

    Article Title: Using antibodies against P2Y and P2X receptors in purinergic signaling research

    Journal: Purinergic Signalling

    doi: 10.1007/s11302-011-9278-z

    Expression of P2Y receptors in osteoblasts. Using flow cytometry analysis, P2Y 1 , P2Y 2 , P2Y 4 , P2Y 6 , P2Y 11 , P2Y 12 , and P2Y 13 receptors were detected in osteoblasts using their respective Alomone Labs antibodies ( open histograms ). Incubating the antibody with the control peptide antigen was used as a negative control ( filled histograms ). Adapted from reference 41 with permission of Elsevier
    Figure Legend Snippet: Expression of P2Y receptors in osteoblasts. Using flow cytometry analysis, P2Y 1 , P2Y 2 , P2Y 4 , P2Y 6 , P2Y 11 , P2Y 12 , and P2Y 13 receptors were detected in osteoblasts using their respective Alomone Labs antibodies ( open histograms ). Incubating the antibody with the control peptide antigen was used as a negative control ( filled histograms ). Adapted from reference 41 with permission of Elsevier

    Techniques Used: Expressing, Flow Cytometry, Cytometry, Negative Control

    Expression of P2Y receptors in Müller cells of the adult rat and mouse retinal slices. Immunohistochemical staining was accomplished using anti-P2Y 2 receptor (#APR-010) and anti-P2Y 6 receptor (#APR-011) antibodies. a No distinct immunoreactivity for the P2Y 2 receptor ( red ) was observed in Müller cells. However, ganglion cells ( asterisk ) and structures within the inner plexiform layer ( IPL ), inner nuclear layer ( INL ), and outer plexiform layer ( OPL ) display faint staining. b P2Y 4 receptor ( red ) is highly expressed in Müller cell end-feet ( arrows ), as detected using anti-P2Y 4 receptor antibody (#APR-006). Additionally, a band of globular structures, probably representing synapse of bipolar cells in the ON-sublamina ( ON ) of the IPL, is stained for this receptor. c The staining pattern of the P2Y 6 receptor ( red ) is similar to that of P2Y 2 . The insets in ( b ) and ( c ) show the ganglion cell layer ( GCL ) and the INL, respectively, at higher magnification. ONL outer nuclear layer, OFF OFF-sublamina of the IPL, PRS photoreceptor segments. Scale bars , 20 μm; inset scale bars , 10 μm. d Immunohistochemical staining was carried out using anti-P2Y 4 receptor antibody. The slices were co-stained against the glial cell marker cellular retinaldehyde-binding protein ( CRALBP ); co-labeling yielded a yellow-orange merge signal. Arrows , Müller cell end-feet; arrowheads , Müller cell somata. GCL ganglion cell layer, IPL inner plexiform layer, NFL nerve fiber layer, ONL outer nuclear layer, PRS , photoreceptor segments. Scale bars , 20 μm. a–c Adapted from reference 12 with permission of John Wiley and Sons. d Adapted from reference 13 with permission of Wiley-Blackwell
    Figure Legend Snippet: Expression of P2Y receptors in Müller cells of the adult rat and mouse retinal slices. Immunohistochemical staining was accomplished using anti-P2Y 2 receptor (#APR-010) and anti-P2Y 6 receptor (#APR-011) antibodies. a No distinct immunoreactivity for the P2Y 2 receptor ( red ) was observed in Müller cells. However, ganglion cells ( asterisk ) and structures within the inner plexiform layer ( IPL ), inner nuclear layer ( INL ), and outer plexiform layer ( OPL ) display faint staining. b P2Y 4 receptor ( red ) is highly expressed in Müller cell end-feet ( arrows ), as detected using anti-P2Y 4 receptor antibody (#APR-006). Additionally, a band of globular structures, probably representing synapse of bipolar cells in the ON-sublamina ( ON ) of the IPL, is stained for this receptor. c The staining pattern of the P2Y 6 receptor ( red ) is similar to that of P2Y 2 . The insets in ( b ) and ( c ) show the ganglion cell layer ( GCL ) and the INL, respectively, at higher magnification. ONL outer nuclear layer, OFF OFF-sublamina of the IPL, PRS photoreceptor segments. Scale bars , 20 μm; inset scale bars , 10 μm. d Immunohistochemical staining was carried out using anti-P2Y 4 receptor antibody. The slices were co-stained against the glial cell marker cellular retinaldehyde-binding protein ( CRALBP ); co-labeling yielded a yellow-orange merge signal. Arrows , Müller cell end-feet; arrowheads , Müller cell somata. GCL ganglion cell layer, IPL inner plexiform layer, NFL nerve fiber layer, ONL outer nuclear layer, PRS , photoreceptor segments. Scale bars , 20 μm. a–c Adapted from reference 12 with permission of John Wiley and Sons. d Adapted from reference 13 with permission of Wiley-Blackwell

    Techniques Used: Expressing, Immunohistochemistry, Staining, Marker, Binding Assay, Labeling

    Differential expression of P2Y receptors in glial following retinal detachment. Immunohistochemical staining of control retinas and from detached areas of porcine retinas 7 days after surgery was done using anti-P2Y 1 receptor (#APR-009), anti-P2Y 2 receptor (#APR-010), and anti-P2Y 4 receptor (#APR-006) antibodies ( red ). Immunoreactivities for P2Y 1 , P2Y 2 , and P2Y 4 receptor proteins ( red ) coincide with that of intermediate filament constituents vimentin ( green and yellow for merging signal). Arrows , vimentin-positive fibers in the outer retina that were also stained for P2Y 2 protein. GCL ganglion cell layer, INL inner nuclear layer, IPL inner plexiform layer, NFL nerve fiber layer, ONL outer nuclear layer. Scale bars , 20 μm. Adapted from reference 15 with permission of Association for Research in Vision and Ophthalmology
    Figure Legend Snippet: Differential expression of P2Y receptors in glial following retinal detachment. Immunohistochemical staining of control retinas and from detached areas of porcine retinas 7 days after surgery was done using anti-P2Y 1 receptor (#APR-009), anti-P2Y 2 receptor (#APR-010), and anti-P2Y 4 receptor (#APR-006) antibodies ( red ). Immunoreactivities for P2Y 1 , P2Y 2 , and P2Y 4 receptor proteins ( red ) coincide with that of intermediate filament constituents vimentin ( green and yellow for merging signal). Arrows , vimentin-positive fibers in the outer retina that were also stained for P2Y 2 protein. GCL ganglion cell layer, INL inner nuclear layer, IPL inner plexiform layer, NFL nerve fiber layer, ONL outer nuclear layer. Scale bars , 20 μm. Adapted from reference 15 with permission of Association for Research in Vision and Ophthalmology

    Techniques Used: Expressing, Immunohistochemistry, Staining

    Expression of P2Y receptors in rat duodenal epithelium. Immunohistochemical staining was done using anti-P2Y 1 receptor antibody (#APR-009), anti-P2Y 2 receptor antibody (#APR-010), and anti-P2Y 6 receptor antibody (#APR-011). a P2Y 1 is expressed along the brush border membranes of villous cells. P2Y 2 and P2Y 4 receptors are not expressed ( b , c ). d P2Y 6 receptor is diffusely expressed in the villous cells and interstitium. Adapted from reference 31 with permission of John Wiley and Sons
    Figure Legend Snippet: Expression of P2Y receptors in rat duodenal epithelium. Immunohistochemical staining was done using anti-P2Y 1 receptor antibody (#APR-009), anti-P2Y 2 receptor antibody (#APR-010), and anti-P2Y 6 receptor antibody (#APR-011). a P2Y 1 is expressed along the brush border membranes of villous cells. P2Y 2 and P2Y 4 receptors are not expressed ( b , c ). d P2Y 6 receptor is diffusely expressed in the villous cells and interstitium. Adapted from reference 31 with permission of John Wiley and Sons

    Techniques Used: Expressing, Immunohistochemistry, Staining

    Expression of P2Y receptors in rat distal colon. All P2Y immunohistochemical staining was done using Alomone Labs anti-P2Y receptor antibodies. a Double immunolabeling reveals that P2Y 1 is expressed in enteric smooth muscle cells and S100 in enteric glial cells. b In the myenteric plexus, P2Y 2 is detected in neurons. c P2Y 4 is detected in enteric glial cells. d P2Y 6 is expressed in vascular and enteric smooth muscle cells. e P2Y 11 is detected in enteric glial cells and P2Y 12 is expressed in enteric nerves ( f ). Adapted from reference 30 with permission of Elsevier
    Figure Legend Snippet: Expression of P2Y receptors in rat distal colon. All P2Y immunohistochemical staining was done using Alomone Labs anti-P2Y receptor antibodies. a Double immunolabeling reveals that P2Y 1 is expressed in enteric smooth muscle cells and S100 in enteric glial cells. b In the myenteric plexus, P2Y 2 is detected in neurons. c P2Y 4 is detected in enteric glial cells. d P2Y 6 is expressed in vascular and enteric smooth muscle cells. e P2Y 11 is detected in enteric glial cells and P2Y 12 is expressed in enteric nerves ( f ). Adapted from reference 30 with permission of Elsevier

    Techniques Used: Expressing, Immunohistochemistry, Staining, Immunolabeling

    2) Product Images from "Using antibodies against P2Y and P2X receptors in purinergic signaling research"

    Article Title: Using antibodies against P2Y and P2X receptors in purinergic signaling research

    Journal: Purinergic Signalling

    doi: 10.1007/s11302-011-9278-z

    Expression of P2Y receptors in osteoblasts. Using flow cytometry analysis, P2Y 1 , P2Y 2 , P2Y 4 , P2Y 6 , P2Y 11 , P2Y 12 , and P2Y 13 receptors were detected in osteoblasts using their respective Alomone Labs antibodies ( open histograms ). Incubating the antibody with the control peptide antigen was used as a negative control ( filled histograms ). Adapted from reference 41 with permission of Elsevier
    Figure Legend Snippet: Expression of P2Y receptors in osteoblasts. Using flow cytometry analysis, P2Y 1 , P2Y 2 , P2Y 4 , P2Y 6 , P2Y 11 , P2Y 12 , and P2Y 13 receptors were detected in osteoblasts using their respective Alomone Labs antibodies ( open histograms ). Incubating the antibody with the control peptide antigen was used as a negative control ( filled histograms ). Adapted from reference 41 with permission of Elsevier

    Techniques Used: Expressing, Flow Cytometry, Cytometry, Negative Control

    Expression of P2Y receptors in Müller cells of the adult rat and mouse retinal slices. Immunohistochemical staining was accomplished using anti-P2Y 2 receptor (#APR-010) and anti-P2Y 6 receptor (#APR-011) antibodies. a No distinct immunoreactivity for the P2Y 2 receptor ( red ) was observed in Müller cells. However, ganglion cells ( asterisk ) and structures within the inner plexiform layer ( IPL ), inner nuclear layer ( INL ), and outer plexiform layer ( OPL ) display faint staining. b P2Y 4 receptor ( red ) is highly expressed in Müller cell end-feet ( arrows ), as detected using anti-P2Y 4 receptor antibody (#APR-006). Additionally, a band of globular structures, probably representing synapse of bipolar cells in the ON-sublamina ( ON ) of the IPL, is stained for this receptor. c The staining pattern of the P2Y 6 receptor ( red ) is similar to that of P2Y 2 . The insets in ( b ) and ( c ) show the ganglion cell layer ( GCL ) and the INL, respectively, at higher magnification. ONL outer nuclear layer, OFF OFF-sublamina of the IPL, PRS photoreceptor segments. Scale bars , 20 μm; inset scale bars , 10 μm. d Immunohistochemical staining was carried out using anti-P2Y 4 receptor antibody. The slices were co-stained against the glial cell marker cellular retinaldehyde-binding protein ( CRALBP ); co-labeling yielded a yellow-orange merge signal. Arrows , Müller cell end-feet; arrowheads , Müller cell somata. GCL ganglion cell layer, IPL inner plexiform layer, NFL nerve fiber layer, ONL outer nuclear layer, PRS , photoreceptor segments. Scale bars , 20 μm. a–c Adapted from reference 12 with permission of John Wiley and Sons. d Adapted from reference 13 with permission of Wiley-Blackwell
    Figure Legend Snippet: Expression of P2Y receptors in Müller cells of the adult rat and mouse retinal slices. Immunohistochemical staining was accomplished using anti-P2Y 2 receptor (#APR-010) and anti-P2Y 6 receptor (#APR-011) antibodies. a No distinct immunoreactivity for the P2Y 2 receptor ( red ) was observed in Müller cells. However, ganglion cells ( asterisk ) and structures within the inner plexiform layer ( IPL ), inner nuclear layer ( INL ), and outer plexiform layer ( OPL ) display faint staining. b P2Y 4 receptor ( red ) is highly expressed in Müller cell end-feet ( arrows ), as detected using anti-P2Y 4 receptor antibody (#APR-006). Additionally, a band of globular structures, probably representing synapse of bipolar cells in the ON-sublamina ( ON ) of the IPL, is stained for this receptor. c The staining pattern of the P2Y 6 receptor ( red ) is similar to that of P2Y 2 . The insets in ( b ) and ( c ) show the ganglion cell layer ( GCL ) and the INL, respectively, at higher magnification. ONL outer nuclear layer, OFF OFF-sublamina of the IPL, PRS photoreceptor segments. Scale bars , 20 μm; inset scale bars , 10 μm. d Immunohistochemical staining was carried out using anti-P2Y 4 receptor antibody. The slices were co-stained against the glial cell marker cellular retinaldehyde-binding protein ( CRALBP ); co-labeling yielded a yellow-orange merge signal. Arrows , Müller cell end-feet; arrowheads , Müller cell somata. GCL ganglion cell layer, IPL inner plexiform layer, NFL nerve fiber layer, ONL outer nuclear layer, PRS , photoreceptor segments. Scale bars , 20 μm. a–c Adapted from reference 12 with permission of John Wiley and Sons. d Adapted from reference 13 with permission of Wiley-Blackwell

    Techniques Used: Expressing, Immunohistochemistry, Staining, Marker, Binding Assay, Labeling

    Differential expression of P2Y receptors in glial following retinal detachment. Immunohistochemical staining of control retinas and from detached areas of porcine retinas 7 days after surgery was done using anti-P2Y 1 receptor (#APR-009), anti-P2Y 2 receptor (#APR-010), and anti-P2Y 4 receptor (#APR-006) antibodies ( red ). Immunoreactivities for P2Y 1 , P2Y 2 , and P2Y 4 receptor proteins ( red ) coincide with that of intermediate filament constituents vimentin ( green and yellow for merging signal). Arrows , vimentin-positive fibers in the outer retina that were also stained for P2Y 2 protein. GCL ganglion cell layer, INL inner nuclear layer, IPL inner plexiform layer, NFL nerve fiber layer, ONL outer nuclear layer. Scale bars , 20 μm. Adapted from reference 15 with permission of Association for Research in Vision and Ophthalmology
    Figure Legend Snippet: Differential expression of P2Y receptors in glial following retinal detachment. Immunohistochemical staining of control retinas and from detached areas of porcine retinas 7 days after surgery was done using anti-P2Y 1 receptor (#APR-009), anti-P2Y 2 receptor (#APR-010), and anti-P2Y 4 receptor (#APR-006) antibodies ( red ). Immunoreactivities for P2Y 1 , P2Y 2 , and P2Y 4 receptor proteins ( red ) coincide with that of intermediate filament constituents vimentin ( green and yellow for merging signal). Arrows , vimentin-positive fibers in the outer retina that were also stained for P2Y 2 protein. GCL ganglion cell layer, INL inner nuclear layer, IPL inner plexiform layer, NFL nerve fiber layer, ONL outer nuclear layer. Scale bars , 20 μm. Adapted from reference 15 with permission of Association for Research in Vision and Ophthalmology

    Techniques Used: Expressing, Immunohistochemistry, Staining

    Expression of P2Y receptors in rat duodenal epithelium. Immunohistochemical staining was done using anti-P2Y 1 receptor antibody (#APR-009), anti-P2Y 2 receptor antibody (#APR-010), and anti-P2Y 6 receptor antibody (#APR-011). a P2Y 1 is expressed along the brush border membranes of villous cells. P2Y 2 and P2Y 4 receptors are not expressed ( b , c ). d P2Y 6 receptor is diffusely expressed in the villous cells and interstitium. Adapted from reference 31 with permission of John Wiley and Sons
    Figure Legend Snippet: Expression of P2Y receptors in rat duodenal epithelium. Immunohistochemical staining was done using anti-P2Y 1 receptor antibody (#APR-009), anti-P2Y 2 receptor antibody (#APR-010), and anti-P2Y 6 receptor antibody (#APR-011). a P2Y 1 is expressed along the brush border membranes of villous cells. P2Y 2 and P2Y 4 receptors are not expressed ( b , c ). d P2Y 6 receptor is diffusely expressed in the villous cells and interstitium. Adapted from reference 31 with permission of John Wiley and Sons

    Techniques Used: Expressing, Immunohistochemistry, Staining

    Expression of P2Y receptors in rat distal colon. All P2Y immunohistochemical staining was done using Alomone Labs anti-P2Y receptor antibodies. a Double immunolabeling reveals that P2Y 1 is expressed in enteric smooth muscle cells and S100 in enteric glial cells. b In the myenteric plexus, P2Y 2 is detected in neurons. c P2Y 4 is detected in enteric glial cells. d P2Y 6 is expressed in vascular and enteric smooth muscle cells. e P2Y 11 is detected in enteric glial cells and P2Y 12 is expressed in enteric nerves ( f ). Adapted from reference 30 with permission of Elsevier
    Figure Legend Snippet: Expression of P2Y receptors in rat distal colon. All P2Y immunohistochemical staining was done using Alomone Labs anti-P2Y receptor antibodies. a Double immunolabeling reveals that P2Y 1 is expressed in enteric smooth muscle cells and S100 in enteric glial cells. b In the myenteric plexus, P2Y 2 is detected in neurons. c P2Y 4 is detected in enteric glial cells. d P2Y 6 is expressed in vascular and enteric smooth muscle cells. e P2Y 11 is detected in enteric glial cells and P2Y 12 is expressed in enteric nerves ( f ). Adapted from reference 30 with permission of Elsevier

    Techniques Used: Expressing, Immunohistochemistry, Staining, Immunolabeling

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    Alomone Labs anti p2y2 receptor antibody
    The RGD motif in P2Y 2 is required for extracellular ATP-driven cancer cell invasion. A Schematic diagram of the hanging drop sphere model for 3D sphere invasion assays. B Brightfield and fluorescent images of spheres formed using AsPC-1 cells (magenta) with a histone 2B (H2B) tagged with red fluorescent protein (RFP) and the stellate cell line PS-1 (green) with H2B tagged with a green fluorescent protein (GFP). Middle pannel shows AsPC-1 cells in spheres with a dotted line highlighting the central sphere area. Spheres were treated with vehicle control or 100 μM ATP alone or with 5 μM AR-C or 10 μM cRGDfV. The quantification is shown in C using SuperPlots, where each colour represents a repeat and the larger points represent the mean % Invasion for each repeat. D Quantification of spheres formed by AsPC-1 cells transfected with a control siRNA or P2Y 2 siRNA and treated with or without 100 μM ATP. E Brightfield and fluorescent images of spheres formed by AsPC-1 cells subjected to CRISPR/Cas9 gene disruption using a control guide RNA (CTR CRISPR ) or P2Y 2 guide RNAs (P2Y 2 CRISPR ) and treated with or without 100 μM ATP. Quantification in F . G, I Brightfield and fluorescent images of AsPC-1 P2Y 2 CRISPR cells or PANC-1 cells (respectively) transfected with wild-type <t>P2RY2</t> (P2Y 2 RGD ) or mutant P2RY2 D97E (P2Y 2 RGE ) treated with or without 100 μM ATP and its quantification in H and J , respectively. Statistical analysis with Kuskal-Wallis multiple comparison test.
    Anti P2y2 Receptor Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p2y2 receptor antibody/product/Alomone Labs
    Average 94 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    anti p2y2 receptor antibody - by Bioz Stars, 2022-12
    94/100 stars
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    94
    Alomone Labs anti p2y1 receptor antibody
    Extracellular adenosine triphosphate (ATP) prevents glutamate-induced excitotoxicity via binding <t>P2Y1</t> receptors in SH-SY5Y cells. (A) Representative images of the cell morphology after cultured SH-SY5Y cells were treated with various concentrations of glutamate (Glu) for 24 h; the control group (Ctrl) consisted of the untreated cells. Scale bar = 100 μm under 20× magnification. (B) Cell viability was detected using Cell Counting kit (CCK-8). (C) Representative images of the cell morphology after pretreated by ATP (5 and 10 μM) or phosphate-buffered saline (PBS) following by the application of 10 mM glutamate for 24 h. Scale bar = 100 μm under 20× magnification. (D) Cell viability was calculated. (E) The protein expressions of P2Y1 <t>receptor,</t> and the bar chart presented the levels of P2Y1 receptor in all groups. Expression of GAPDH was served as a loading control. (F) CCK-8 kit was performed to evaluated cell viability after the administration of MRS2500, and the changes were shown in histograms as percentage with control. All data represent mean ± SEM from at least three independent experiments. ** P
    Anti P2y1 Receptor 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 p2y1 receptor antibody/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti p2y1 receptor antibody - by Bioz Stars, 2022-12
    94/100 stars
      Buy from Supplier

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    The RGD motif in P2Y 2 is required for extracellular ATP-driven cancer cell invasion. A Schematic diagram of the hanging drop sphere model for 3D sphere invasion assays. B Brightfield and fluorescent images of spheres formed using AsPC-1 cells (magenta) with a histone 2B (H2B) tagged with red fluorescent protein (RFP) and the stellate cell line PS-1 (green) with H2B tagged with a green fluorescent protein (GFP). Middle pannel shows AsPC-1 cells in spheres with a dotted line highlighting the central sphere area. Spheres were treated with vehicle control or 100 μM ATP alone or with 5 μM AR-C or 10 μM cRGDfV. The quantification is shown in C using SuperPlots, where each colour represents a repeat and the larger points represent the mean % Invasion for each repeat. D Quantification of spheres formed by AsPC-1 cells transfected with a control siRNA or P2Y 2 siRNA and treated with or without 100 μM ATP. E Brightfield and fluorescent images of spheres formed by AsPC-1 cells subjected to CRISPR/Cas9 gene disruption using a control guide RNA (CTR CRISPR ) or P2Y 2 guide RNAs (P2Y 2 CRISPR ) and treated with or without 100 μM ATP. Quantification in F . G, I Brightfield and fluorescent images of AsPC-1 P2Y 2 CRISPR cells or PANC-1 cells (respectively) transfected with wild-type P2RY2 (P2Y 2 RGD ) or mutant P2RY2 D97E (P2Y 2 RGE ) treated with or without 100 μM ATP and its quantification in H and J , respectively. Statistical analysis with Kuskal-Wallis multiple comparison test.

    Journal: bioRxiv

    Article Title: Extracellular ATP drives pancreatic cancer cell invasion via purinergic receptor-integrin interactions

    doi: 10.1101/2022.10.24.513477

    Figure Lengend Snippet: The RGD motif in P2Y 2 is required for extracellular ATP-driven cancer cell invasion. A Schematic diagram of the hanging drop sphere model for 3D sphere invasion assays. B Brightfield and fluorescent images of spheres formed using AsPC-1 cells (magenta) with a histone 2B (H2B) tagged with red fluorescent protein (RFP) and the stellate cell line PS-1 (green) with H2B tagged with a green fluorescent protein (GFP). Middle pannel shows AsPC-1 cells in spheres with a dotted line highlighting the central sphere area. Spheres were treated with vehicle control or 100 μM ATP alone or with 5 μM AR-C or 10 μM cRGDfV. The quantification is shown in C using SuperPlots, where each colour represents a repeat and the larger points represent the mean % Invasion for each repeat. D Quantification of spheres formed by AsPC-1 cells transfected with a control siRNA or P2Y 2 siRNA and treated with or without 100 μM ATP. E Brightfield and fluorescent images of spheres formed by AsPC-1 cells subjected to CRISPR/Cas9 gene disruption using a control guide RNA (CTR CRISPR ) or P2Y 2 guide RNAs (P2Y 2 CRISPR ) and treated with or without 100 μM ATP. Quantification in F . G, I Brightfield and fluorescent images of AsPC-1 P2Y 2 CRISPR cells or PANC-1 cells (respectively) transfected with wild-type P2RY2 (P2Y 2 RGD ) or mutant P2RY2 D97E (P2Y 2 RGE ) treated with or without 100 μM ATP and its quantification in H and J , respectively. Statistical analysis with Kuskal-Wallis multiple comparison test.

    Article Snippet: DNA-antibody coupling reactionDNA labelling of anti-aV antibody (P2W7, Santa Cruz) and anti-P2Y2 receptor antibody (APR-010, Alomone labs) was performed via maleimidePEG2-succinimidyl ester coupling reaction as previously described ( ; ).

    Techniques: Transfection, CRISPR, Mutagenesis

    Extracellular adenosine triphosphate (ATP) prevents glutamate-induced excitotoxicity via binding P2Y1 receptors in SH-SY5Y cells. (A) Representative images of the cell morphology after cultured SH-SY5Y cells were treated with various concentrations of glutamate (Glu) for 24 h; the control group (Ctrl) consisted of the untreated cells. Scale bar = 100 μm under 20× magnification. (B) Cell viability was detected using Cell Counting kit (CCK-8). (C) Representative images of the cell morphology after pretreated by ATP (5 and 10 μM) or phosphate-buffered saline (PBS) following by the application of 10 mM glutamate for 24 h. Scale bar = 100 μm under 20× magnification. (D) Cell viability was calculated. (E) The protein expressions of P2Y1 receptor, and the bar chart presented the levels of P2Y1 receptor in all groups. Expression of GAPDH was served as a loading control. (F) CCK-8 kit was performed to evaluated cell viability after the administration of MRS2500, and the changes were shown in histograms as percentage with control. All data represent mean ± SEM from at least three independent experiments. ** P

    Journal: Frontiers in Neuroscience

    Article Title: Extracellular Adenosine Triphosphate Binding to P2Y1 Receptors Prevents Glutamate-Induced Excitotoxicity: Involvement of Erk1/2 Signaling Pathway to Suppress Autophagy

    doi: 10.3389/fnins.2022.901688

    Figure Lengend Snippet: Extracellular adenosine triphosphate (ATP) prevents glutamate-induced excitotoxicity via binding P2Y1 receptors in SH-SY5Y cells. (A) Representative images of the cell morphology after cultured SH-SY5Y cells were treated with various concentrations of glutamate (Glu) for 24 h; the control group (Ctrl) consisted of the untreated cells. Scale bar = 100 μm under 20× magnification. (B) Cell viability was detected using Cell Counting kit (CCK-8). (C) Representative images of the cell morphology after pretreated by ATP (5 and 10 μM) or phosphate-buffered saline (PBS) following by the application of 10 mM glutamate for 24 h. Scale bar = 100 μm under 20× magnification. (D) Cell viability was calculated. (E) The protein expressions of P2Y1 receptor, and the bar chart presented the levels of P2Y1 receptor in all groups. Expression of GAPDH was served as a loading control. (F) CCK-8 kit was performed to evaluated cell viability after the administration of MRS2500, and the changes were shown in histograms as percentage with control. All data represent mean ± SEM from at least three independent experiments. ** P

    Article Snippet: Anti-P2Y1 receptor antibody was purchased from Alomone Labs (Jerusalem BioPark, Israel).

    Techniques: Binding Assay, Cell Culture, Cell Counting, CCK-8 Assay, Expressing