anti rabbit p2y 6 r polyclonal antibody  (Alomone Labs)

Journal: Purinergic Signalling

Article Title: Proteasomal-dependent endothelial P2Y 6 receptor downregulation as an adaptive mechanism limiting monocyte adhesion during intestinal schistosomiasis

doi: 10.1007/s11302-026-10141-x

Figure Lengend Snippet: Endothelial P2Y 6 R activation failed to increase monocyte adhesion during schistosomiasis. A. Monocyte adhesion induced by UDP 100 µM was blocked by the selective P2Y 6 R antagonist (MRS2578 1 µM) in the control group (white bars). B. UDP 100 µM stimulus did not increase monocyte adhesion in the infected group (gray bars). ARL67156 100 µM = ectonucleotidase inhibitor (added 30 min before). Data were expressed as mean and SEM of n independent experiments performed in triplicate for each condition. ***p < 0.001 (One-way ANOVA followed by Tukey’s multiple comparisons test, n = 4 animals for each group)

Article Snippet: After that, cells were washed twice for 5 min with Triton X-100 0.2% diluted in PBS and incubated with anti-rabbit P2Y 6 R polyclonal antibody (1:500, Alomone Labs, Israel) overnight.

Techniques: Activation Assay, Control, Infection

Journal: Purinergic Signalling

Article Title: Proteasomal-dependent endothelial P2Y 6 receptor downregulation as an adaptive mechanism limiting monocyte adhesion during intestinal schistosomiasis

doi: 10.1007/s11302-026-10141-x

Figure Lengend Snippet: Endothelial P2Y 6 R mRNA and protein levels are reduced during schistosomiasis. A. Immunocytochemistry staining of cultured mesenteric endothelial cells from control (top line) and infected (bottom line) groups using anti-P2Y 6 R antibody (1:500, overnight), followed by Alexa Fluor 488 secondary antibody (1:300, 1 h, green) and nuclear fluorescence using DAPI (blue) (scale bar = 20 μm; × 400) (see methods). B. The fluorescence intensity was quantified and then expressed as arbitrary units (a.u.). P2Y 6 R mRNA levels were lower in the infected (gray bars) than in the control (white bars) group. C. The P2Y 6 R mRNA levels were normalized by the endogenous gene β-actin. Data were expressed as mean and SEM of n independent cultures performed with different animals for each condition. ***p < 0.001, **p < 0.01 (unpaired t -test, n = 3–4 animals for each group)

Article Snippet: After that, cells were washed twice for 5 min with Triton X-100 0.2% diluted in PBS and incubated with anti-rabbit P2Y 6 R polyclonal antibody (1:500, Alomone Labs, Israel) overnight.

Techniques: Immunocytochemistry, Staining, Cell Culture, Control, Infection, Fluorescence

Journal: Purinergic Signalling

Article Title: Proteasomal-dependent endothelial P2Y 6 receptor downregulation as an adaptive mechanism limiting monocyte adhesion during intestinal schistosomiasis

doi: 10.1007/s11302-026-10141-x

Figure Lengend Snippet: Endothelial P2Y 6 R downregulation is mediated by proteosome degradation during schistosomiasis. A. Treatment with the ectonucleotidase inhibitor ( ARL67156 100 µM, 24 h) followed by a wash step and UDP 100 µM (5 h); treatment with apyrase 2 U/L (24 h) followed by washing and UDP 100 µM (5 h). B. Treatment with the proteosome inhibitor MG132 (1 µM, 1 h) followed by UDP (100 µM, 5 h). Data were expressed as mean and SEM of n independent cultures in triplicate for each condition. ***p < 0.001, *p < 0.05 (One-way ANOVA followed by Tukey’s multiple comparisons test, n = 3 animals for each group). ARL = ARL67156

Article Snippet: After that, cells were washed twice for 5 min with Triton X-100 0.2% diluted in PBS and incubated with anti-rabbit P2Y 6 R polyclonal antibody (1:500, Alomone Labs, Israel) overnight.

Techniques:

Journal: Purinergic Signalling

Article Title: Proteasomal-dependent endothelial P2Y 6 receptor downregulation as an adaptive mechanism limiting monocyte adhesion during intestinal schistosomiasis

doi: 10.1007/s11302-026-10141-x

Figure Lengend Snippet: Endothelial P2Y 6 R expression is increased by proteosome inhibition during schistosomiasis. A. Immunocytochemistry staining of cultured mesenteric endothelial cells from basal (top line) MG132 1 µM, 1 h (middle line) or MG132 1 µM, 6 h (bottom line) using anti-P2Y 6 R antibody (1:500, overnight) followed by Alexa Fluor 488 secondary antibody secondary antibody (1:300, 1 h, green) and nuclear fluorescence using DAPI (blue) (scale bar = 20 μm; × 400) (see methods). B. The fluorescence intensity was quantified for all groups and then expressed as arbitrary units (a.u.). C . ROS was quantified colorimetrically using NBT. D . Lipid peroxidation was quantified by the production of malondialdehyde (MDA) in reaction with thiobarbituric acid. Data were expressed as the mean and SEM of n independent cultures in triplicate for each condition. **p < 0.01 ( B , One-way ANOVA followed by Tukey’s multiple comparisons test, n = 3 animals for each group); ***p < 0.001; **p < 0.01 (C, D, unpaired t -test, n = 3 animals for each group). ROS = reactive oxygen species; TBARS = thiobarbituric acid reactive species

Article Snippet: After that, cells were washed twice for 5 min with Triton X-100 0.2% diluted in PBS and incubated with anti-rabbit P2Y 6 R polyclonal antibody (1:500, Alomone Labs, Israel) overnight.

Techniques: Expressing, Inhibition, Immunocytochemistry, Staining, Cell Culture, Fluorescence

Journal: Purinergic Signalling

Article Title: Proteasomal-dependent endothelial P2Y 6 receptor downregulation as an adaptive mechanism limiting monocyte adhesion during intestinal schistosomiasis

doi: 10.1007/s11302-026-10141-x

Figure Lengend Snippet: Proposed mechanism of proteasomal-dependent P2Y 6 receptor downregulation pathway in mesenteric endothelial cells in a preclinical model of intestinal schistosomiasis. The mesenteric endothelial cells primed by infection showed a proteasome-dependent P2Y 6 receptor downregulation under conditions of oxidative stress (1) triggering post-translational modifications protein such as polyubiquitination (2), which targets the receptor to the proteasomal degradation pathway (3–4), ultimately resulting in reduced endothelial P2Y 6 R expression and diminished proinflammatory effect (5)

Article Snippet: After that, cells were washed twice for 5 min with Triton X-100 0.2% diluted in PBS and incubated with anti-rabbit P2Y 6 R polyclonal antibody (1:500, Alomone Labs, Israel) overnight.

Techniques: Infection, Expressing

Journal: Purinergic Signalling

Article Title: Proteasomal-dependent endothelial P2Y 6 receptor downregulation as an adaptive mechanism limiting monocyte adhesion during intestinal schistosomiasis

doi: 10.1007/s11302-026-10141-x

Figure Lengend Snippet: Endothelial P2Y 6 R mRNA and protein levels are reduced during schistosomiasis. A. Immunocytochemistry staining of cultured mesenteric endothelial cells from control (top line) and infected (bottom line) groups using anti-P2Y 6 R antibody (1:500, overnight), followed by Alexa Fluor 488 secondary antibody (1:300, 1 h, green) and nuclear fluorescence using DAPI (blue) (scale bar = 20 μm; × 400) (see methods). B. The fluorescence intensity was quantified and then expressed as arbitrary units (a.u.). P2Y 6 R mRNA levels were lower in the infected (gray bars) than in the control (white bars) group. C. The P2Y 6 R mRNA levels were normalized by the endogenous gene β-actin. Data were expressed as mean and SEM of n independent cultures performed with different animals for each condition. ***p < 0.001, **p < 0.01 (unpaired t -test, n = 3–4 animals for each group)

Article Snippet: Primary antibody anti-P2Y 6 R (#APR-011) was obtained from Alomone Labs.

Techniques: Immunocytochemistry, Staining, Cell Culture, Control, Infection, Fluorescence

Journal: Purinergic Signalling

Article Title: Proteasomal-dependent endothelial P2Y 6 receptor downregulation as an adaptive mechanism limiting monocyte adhesion during intestinal schistosomiasis

doi: 10.1007/s11302-026-10141-x

Figure Lengend Snippet: Endothelial P2Y 6 R expression is increased by proteosome inhibition during schistosomiasis. A. Immunocytochemistry staining of cultured mesenteric endothelial cells from basal (top line) MG132 1 µM, 1 h (middle line) or MG132 1 µM, 6 h (bottom line) using anti-P2Y 6 R antibody (1:500, overnight) followed by Alexa Fluor 488 secondary antibody secondary antibody (1:300, 1 h, green) and nuclear fluorescence using DAPI (blue) (scale bar = 20 μm; × 400) (see methods). B. The fluorescence intensity was quantified for all groups and then expressed as arbitrary units (a.u.). C . ROS was quantified colorimetrically using NBT. D . Lipid peroxidation was quantified by the production of malondialdehyde (MDA) in reaction with thiobarbituric acid. Data were expressed as the mean and SEM of n independent cultures in triplicate for each condition. **p < 0.01 ( B , One-way ANOVA followed by Tukey’s multiple comparisons test, n = 3 animals for each group); ***p < 0.001; **p < 0.01 (C, D, unpaired t -test, n = 3 animals for each group). ROS = reactive oxygen species; TBARS = thiobarbituric acid reactive species

Article Snippet: Primary antibody anti-P2Y 6 R (#APR-011) was obtained from Alomone Labs.

Techniques: Expressing, Inhibition, Immunocytochemistry, Staining, Cell Culture, Fluorescence

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: Purinergic Signalling

Article Title: Analysis of purine receptor expression and functionality in alveolar epithelial cells

doi: 10.1007/s11302-020-09696-0

Figure Lengend Snippet: siRNA knockdown experiments of selective P2 receptors in L2, R3/1, RLE, and A549 cells. a Relative expression of the P2 receptor genes after siRNA knockdown using real-time quantitative RT-PCR (scr: scramble siRNA; KD: respective siRNA as indicated), n = 3–4 for each receptor and each AEC line investigated, mean ± SD. b [Ca2+]cyt following stimulation with ATP (100 μM) after siRNA knockdown as indicated. Displayed are differences between maximum and baseline [Ca2+]cyt. (∆: delta) as box plots, with median values ± 75% and 25% quartiles, n = 10–31, * p < 0.001 vs. baseline conditions, # p < 0.001 vs ctr

Article Snippet: SiRNA knockdown of P2Rs The cells were transfected with 10 nM siRNA for P2Y 2 -, P2Y 6 -, P2Y 4 -, P2X 4 -, P2X 5 -R (Qiagen), or scrambled siRNA (Qiagen), respectively, using the transfection reagent Lipofectamine RNAiMAX (Life Technologies GmbH) according to the manufacturer’s protocol.

Techniques: Knockdown, Expressing, Quantitative RT-PCR