Journal: PLoS ONE

Article Title: Enhancement of Glucose Uptake in Mouse Skeletal Muscle Cells and Adipocytes by P2Y 6 Receptor Agonists

doi: 10.1371/journal.pone.0116203

Figure Lengend Snippet: Structures of P2Y 6 R ligands: nucleotide agonists.

Article Snippet: P2Y 6 R specific antibody was from Alomone Labs Ltd. (Jerusalem, Israel).

Techniques:

Journal: PLoS ONE

Article Title: Enhancement of Glucose Uptake in Mouse Skeletal Muscle Cells and Adipocytes by P2Y 6 Receptor Agonists

doi: 10.1371/journal.pone.0116203

Figure Lengend Snippet: MRS2957 was applied in the presence or absence of P2Y 6 R antagonist MRS2578 (1 µM). * P <0.05, when compared to controls (n = 3).

Article Snippet: P2Y 6 R specific antibody was from Alomone Labs Ltd. (Jerusalem, Israel).

Techniques:

Journal: PLoS ONE

Article Title: Enhancement of Glucose Uptake in Mouse Skeletal Muscle Cells and Adipocytes by P2Y 6 Receptor Agonists

doi: 10.1371/journal.pone.0116203

Figure Lengend Snippet: C2C12 cells (A) or 3T3-L1 cells (B) were preincubated with inhibitors of MAPK (U0126, 10 µM) and AMPK (Compound C, 10 µM) followed by treatment with P2Y 6 R agonist MRS2957. * P <0.05, when compared to controls, # P <0.05, when compared to MRS2957, 100 nM (n = 3).

Article Snippet: P2Y 6 R specific antibody was from Alomone Labs Ltd. (Jerusalem, Israel).

Techniques:

Journal: Molecular Pain

Article Title: Role of spinal P2Y 6 and P2Y 11 receptors in neuropathic pain in rats: possible involvement of glial cells

doi: 10.1186/1744-8069-10-29

Figure Lengend Snippet: Intrathecal injection of the selective P2Y 6,11 receptor antagonists reverses tactile allodynia. Time course of the antiallodynic effects of the selective P2Y 6 (MRS2578, panel A ) and P2Y 11 (NF340, panel C ) receptor antagonists in rats submitted to spinal nerve injury. Data are expressed as mean ± S.E.M. for 6 animals. Withdrawal threshold was assessed 14 days after spinal nerve injury. Dose–response curves of the antiallodynic effect of MRS2578 (10–100 μM, panel B ) and NF340 (0.3–30 μM, panel D ). Data are expressed as the 50% threshold withdrawal against time curve (AUC). * Significantly different from the vehicle (Veh) group ( p < 0.05), as determined by analysis of variance followed by the Student-Newman-Keuls test. N: Naive.

Article Snippet: After that, they were incubated overnight at 4°C in 5% non-fat dry milk/PBS containing rabbit anti-P2Y 6 (BML-SA586-0050, 1:100, Enzo Life Sciences, Farmingdale, NY), rabbit anti-P2Y 11 (AB9590, 1:500, Millipore, Billerica, MA), mouse anti-Iba-1 (MABN92, 1:500, Millipore, Bellerica, MA) or rabbit anti-GFAP (AB5804, 1:1000, Millipore, Bellerica, MA) antibodies.

Techniques: Injection

Journal: Molecular Pain

Article Title: Role of spinal P2Y 6 and P2Y 11 receptors in neuropathic pain in rats: possible involvement of glial cells

doi: 10.1186/1744-8069-10-29

Figure Lengend Snippet: Spinal nerve injury enhances the expression of P2Y 6,11 receptors. Western blot analysis of the P2Y 6 (panel A ) and P2Y 11 (panel B ) receptors expression in the ipsilateral dorsal spinal cord obtained from naïve and spinal nerve injured (SNL) rats. The primary antibody for P2Y 6 and P2Y 11 were pre-adsorbed with the corresponding antigenic peptide (+P, panel C ). Data were normalized against β-actin and are expressed as the mean ± S.E.M. of 3 independent rats. *Significantly ( p < 0.05) different from the naïve (N) group, as determined by one-way analysis of variance followed by the Student-Newman-Keuls test.

Article Snippet: After that, they were incubated overnight at 4°C in 5% non-fat dry milk/PBS containing rabbit anti-P2Y 6 (BML-SA586-0050, 1:100, Enzo Life Sciences, Farmingdale, NY), rabbit anti-P2Y 11 (AB9590, 1:500, Millipore, Billerica, MA), mouse anti-Iba-1 (MABN92, 1:500, Millipore, Bellerica, MA) or rabbit anti-GFAP (AB5804, 1:1000, Millipore, Bellerica, MA) antibodies.

Techniques: Expressing, Western Blot

Journal: Molecular Pain

Article Title: Role of spinal P2Y 6 and P2Y 11 receptors in neuropathic pain in rats: possible involvement of glial cells

doi: 10.1186/1744-8069-10-29

Figure Lengend Snippet: P2Y 6,11 receptor antagonists prevent enhanced expression of P2Y 6,11 receptors. Effect of the repeated intrathecal administration of MRS2578 (panel A ) and NF340 (panel B ) in spinal nerve injury-induced increase in P2Y 6 and P2Y 11 expression in the ipsilateral dorsal spinal cord. Data were normalized against β-actin and are expressed as the mean ± S.E.M. of 3 independent rats. * Significantly ( p < 0.05) different from the naïve (N) group and # significantly ( p < 0.05) different from the spinal nerve ligated (SNL) group, as determined by one-way analysis of variance followed by the Student-Newman-Keuls test.

Article Snippet: After that, they were incubated overnight at 4°C in 5% non-fat dry milk/PBS containing rabbit anti-P2Y 6 (BML-SA586-0050, 1:100, Enzo Life Sciences, Farmingdale, NY), rabbit anti-P2Y 11 (AB9590, 1:500, Millipore, Billerica, MA), mouse anti-Iba-1 (MABN92, 1:500, Millipore, Bellerica, MA) or rabbit anti-GFAP (AB5804, 1:1000, Millipore, Bellerica, MA) antibodies.

Techniques: Expressing

Journal: Molecular Pain

Article Title: Role of spinal P2Y 6 and P2Y 11 receptors in neuropathic pain in rats: possible involvement of glial cells

doi: 10.1186/1744-8069-10-29

Figure Lengend Snippet: Spinal nerve injury increases the expression of P2Y 6 and P2Y 11 : Blockade by minocycline or fluorocitrate. Effect of the repeated intrathecal administration of minocycline (Mino, panel A and B ) or fluorocitrate (Fluo, panel C and D ) in spinal nerve injury-induced increased expression of P2Y 6 and P2Y 11 receptors in the ipsilateral dorsal spinal cord. Data were normalized against β-actin and are expressed as the mean ± S.E.M. of 3 independent rats. * Significantly ( p < 0.05) different from the naïve (N) group and # significantly ( p < 0.05) different from the spinal nerve ligated (SNL) group, as determined by one-way analysis of variance followed by the Student-Newman-Keuls test.

Article Snippet: After that, they were incubated overnight at 4°C in 5% non-fat dry milk/PBS containing rabbit anti-P2Y 6 (BML-SA586-0050, 1:100, Enzo Life Sciences, Farmingdale, NY), rabbit anti-P2Y 11 (AB9590, 1:500, Millipore, Billerica, MA), mouse anti-Iba-1 (MABN92, 1:500, Millipore, Bellerica, MA) or rabbit anti-GFAP (AB5804, 1:1000, Millipore, Bellerica, MA) antibodies.

Techniques: Expressing

Journal: Physiological Reports

Article Title: Changes in P2Y 6 receptor‐mediated vasoreactivity following focal and global ischemia

doi: 10.14814/phy2.15283

Figure Lengend Snippet: Confocal microscope images demonstrating expression of P2Y 6 (FITC, green) in cross‐sections of middle cerebral arteries. DAPI (4′,6‐Diamidine‐2′‐phenylindole dihydrochloride, a blue nucleus stain, has been added. The tMCAO, Sham, and SAH images were generated by superimposing a DAPI stained image on a FITC stained image. The negative control demonstrates the lack of unspecific antibody binding and the autofluorescence of the lamina elastica interna which separates the intima from the underlying layer of smooth muscle cells

Article Snippet: The cell suspensions were stained overnight at +4°C temperature with primary goat anti‐SM22α (1:100, Abcam) and primary rabbit anti‐P2Y 6 (1:100, Alomone), respectively or goat isotope control IgG (Abcam), rabbit isotope control IgG (Abcam) in blocking buffer (PBS + 5% normal donkey serum) protected from light with rocking, respectively.

Techniques: Microscopy, Expressing, Staining, Generated, Negative Control, Binding Assay

Journal: Physiological Reports

Article Title: Changes in P2Y 6 receptor‐mediated vasoreactivity following focal and global ischemia

doi: 10.14814/phy2.15283

Figure Lengend Snippet: Bar graph with scatter plot demonstrating the expression of P2Y 6 receptors on vascular smooth muscle cells determined by flow cytometry. Data are presented as median and IQR, n = 3–6, * p < 0.05

Article Snippet: The cell suspensions were stained overnight at +4°C temperature with primary goat anti‐SM22α (1:100, Abcam) and primary rabbit anti‐P2Y 6 (1:100, Alomone), respectively or goat isotope control IgG (Abcam), rabbit isotope control IgG (Abcam) in blocking buffer (PBS + 5% normal donkey serum) protected from light with rocking, respectively.

Techniques: Expressing, Flow Cytometry

Journal: Physiological Reports

Article Title: Changes in P2Y 6 receptor‐mediated vasoreactivity following focal and global ischemia

doi: 10.14814/phy2.15283

Figure Lengend Snippet: Representative dot plots and histograms of the experimental setup and analysis of flow cytometry data. (a) Dot plot histogram for entire cell population after isolation. (b) Dot plot histogram of single cells population according to FSC‐H (H—high) versus FSC‐A (A—area scaling). (c) Representative histogram of negative single cells population for Fixable Viability Dye eFluor 780 (viable cells, live/dead); (d) Representative histogram demonstrating SM22α‐positive events of viable cells suspension (log scale). (e) Representative histograms demonstrating P2Y 6 ‐positive events of viable VSMC suspension (log scale)

Article Snippet: The cell suspensions were stained overnight at +4°C temperature with primary goat anti‐SM22α (1:100, Abcam) and primary rabbit anti‐P2Y 6 (1:100, Alomone), respectively or goat isotope control IgG (Abcam), rabbit isotope control IgG (Abcam) in blocking buffer (PBS + 5% normal donkey serum) protected from light with rocking, respectively.

Techniques: Flow Cytometry, Isolation