Journal: PLoS ONE

Article Title: Sensory Dysfunction of Bladder Mucosa and Bladder Oversensitivity in a Rat Model of Metabolic Syndrome

doi: 10.1371/journal.pone.0045578

Figure Lengend Snippet: Western blot analysis with specific antibodies to the TRPV1 receptor, purinergic P2X 2 receptor, and P2X 3 receptor of the rat mucosal layer and the purinergic P2X 1 receptor of the rat smooth muscle layer in controls and FFRs with different in cystometric presentations. A. TRPV1 receptor: The TRPV1 antibody produced a clear single band at 95kDa. B. Purinergic P2X 2 receptor C. Purinergic P2X 3 mature receptor: the predominant P2X 3 form (65kDa). D. Up-regulation of P2X3 native and intermediate polypeptides (up to 55kD) were shown in both FFR groups. E. Purinergic P2X 1 receptor Experiments were repeated two times and representative blots are shown. Data of proteins expression (ratios of signal intensities of investigated receptors relative to β-actin or GAPDH) were calculated with 8 samples in each group. These data of Mean ± SE were standardized and expressed in percentage in which the value of the control group is treated as 100%. Theses values were shown in the bar graph. An asterisk indicates a significant difference between controls and FFR groups (One-way ANOVA with Dunnett’s test, p<0.05).

Article Snippet: Antibodies raised against TRPV1 receptor (Alomone, Israel; 1∶1000 dilution), P2X 2 receptor (Alomone, Israel; 1∶500 dilution), P2X 3 receptor (Neuromics, MN; 1∶500 dilution), P2X 1 receptor (Alomone, Israel; 1∶500 dilution), iNOS (Cayman Chemical, Michigan; 1∶500 dilution), eNOS (BD Biosciences, CA; 1∶1000 dilution), GAPDH (Millipore, CA; 1∶10000 dilution ) and β-actin (Millipore, CA; 1∶10000 dilution) were used.

Techniques: Western Blot, Produced, Expressing

Journal: Poultry Science

Article Title: The release of zearalenone-induced heterophil extracellular traps in chickens is associated with autophagy, glycolysis, PAD enzyme, and P2X 1 receptor

doi: 10.1016/j.psj.2023.102946

Figure Lengend Snippet: The effect of PAD enzyme and P2X 1 receptor in ZEA-triggered HET formation. Heterophils (2 × 10 5 /well) were pretreated with the inhibitor of PAD (Cl-amidine), P2X 1 (NF449) for 30 min, followed by exposure to ZEA (80 μM) for 2 h. Extracellular DNA was quantified by PicoGreen-derived fluorescence intensities using the fluorescence plate reader at 485 nm excitation/525 nm emission. One-way ANOVA was used to determine the significance ( n = 6, **** P < 0.0001).

Article Snippet: The inhibitor of mTOR (Rapamycin, 50 μM), extracellular regulated protein kinases ( ERK ) signaling (U0126, 50 μM), PAD enzyme (Cl-amidine, 6 μM), P2X 1 receptor (NF449, 10 μM), PI3K class I (wortmannin, 50 μM) were obtained from MedChemExpress (New Jersey).

Techniques: Derivative Assay, Fluorescence

Journal: PLoS ONE

Article Title: Sensory Dysfunction of Bladder Mucosa and Bladder Oversensitivity in a Rat Model of Metabolic Syndrome

doi: 10.1371/journal.pone.0045578

Figure Lengend Snippet: Western blot analysis with specific antibodies to the TRPV1 receptor, purinergic P2X 2 receptor, and P2X 3 receptor of the rat mucosal layer and the purinergic P2X 1 receptor of the rat smooth muscle layer in controls and FFRs with different in cystometric presentations. A. TRPV1 receptor: The TRPV1 antibody produced a clear single band at 95kDa. B. Purinergic P2X 2 receptor C. Purinergic P2X 3 mature receptor: the predominant P2X 3 form (65kDa). D. Up-regulation of P2X3 native and intermediate polypeptides (up to 55kD) were shown in both FFR groups. E. Purinergic P2X 1 receptor Experiments were repeated two times and representative blots are shown. Data of proteins expression (ratios of signal intensities of investigated receptors relative to β-actin or GAPDH) were calculated with 8 samples in each group. These data of Mean ± SE were standardized and expressed in percentage in which the value of the control group is treated as 100%. Theses values were shown in the bar graph. An asterisk indicates a significant difference between controls and FFR groups (One-way ANOVA with Dunnett’s test, p<0.05).

Article Snippet: After performing control cystometrographies with saline infusion, rats were subjected to one of four types of physiological experiments: (a) repeatable responses to acidic ATP solution stimulation; (b) capsaicin pretreated to inhibit the response of acidic ATP solution; (c) intravesical administration of liposome (2 mg/ml; Lipella Pharmaceuticals, Pittsburgh, Pennsylvania), a mucosal protective agent , , to partially normalize stimulation by acidic ATP solution; or (d) intravenous administration of pyridoxal 5-phosphate 6-azophenyl-2′,4′-disulfonic acid (PPADS, 70 µmol/kg; Sigma), a commonly used noncompetitively antagonist for heteromeric P2X 2 /P2X 3 , homomeric P2X 3 and P2X 1 receptors , to reverse the stimulation by acidic ATP solution.

Techniques: Western Blot, Produced, Expressing

Journal: BMC Immunology

Article Title: Expression of P2 receptors in human B cells and Epstein-Barr virus-transformed lymphoblastoid cell lines

doi: 10.1186/1471-2172-7-22

Figure Lengend Snippet: Relative expression of P2 receptors in B cells . The relative expression of the P2X receptor gene (upper; P2X 1 , P2X 2 , P2X 3 , P2X 4 , P2X 5 , P2X 6 , and P2X 7 ) and P2Y receptor gene (lower; P2Y 1 , P2Y 2 , P2Y 4 , P2Y 6 , P2Y 11 , P2Y 12 , P2Y 13 , and P2Y 14 ) in B cells are presented (n = 4). The expression was normalized to GAPDH. P2X receptors were calibrated by P2X 1 and P2Y receptors were calibrated by P2Y 1 . Data are the mean ± SEM.

Article Snippet: The membrane was incubated with rabbit polyclonal antibodies (Alomone Labs, Jerusalem, Israel) against P2X 1 receptor, P2X 4 receptor, P2X 7 receptor, P2Y 1 receptor, or P2Y 11 receptor in TBST for 2 hours at room temperature, washed with TBST, and incubated with secondary anti-rabbit IgG (Amersham Pharmacia Biotech) in TBST for 1 hour.

Techniques: Expressing

Journal: BMC Immunology

Article Title: Expression of P2 receptors in human B cells and Epstein-Barr virus-transformed lymphoblastoid cell lines

doi: 10.1186/1471-2172-7-22

Figure Lengend Snippet: Comparison of relative P2 receptor expression in B cells and LCLs . The relative expression of the P2X receptor gene (upper) and P2Y receptor gene (lower) in B cells, EBV-infected B cells for 2 weeks, and LCLs are presented (n = 4). The expression was normalized to GAPDH. P2X receptors were calibrated by P2X 1 of B cells and P2Y receptors were calibrated by P2Y 1 of B cells. Data are the mean ± SEM.

Article Snippet: The membrane was incubated with rabbit polyclonal antibodies (Alomone Labs, Jerusalem, Israel) against P2X 1 receptor, P2X 4 receptor, P2X 7 receptor, P2Y 1 receptor, or P2Y 11 receptor in TBST for 2 hours at room temperature, washed with TBST, and incubated with secondary anti-rabbit IgG (Amersham Pharmacia Biotech) in TBST for 1 hour.

Techniques: Expressing, Infection

Journal: BMC Immunology

Article Title: Expression of P2 receptors in human B cells and Epstein-Barr virus-transformed lymphoblastoid cell lines

doi: 10.1186/1471-2172-7-22

Figure Lengend Snippet: Relative expression of P2 receptors in PBMCs . The relative expression of the P2X receptor gene (upper) and P2Y receptor gene (lower) in PBMCs are presented (n = 4). The expression was normalized to GAPDH. P2X receptors were calibrated by P2X 1 and P2Y receptors were calibrated by P2Y 1 . Data are the mean ± SEM.

Article Snippet: The membrane was incubated with rabbit polyclonal antibodies (Alomone Labs, Jerusalem, Israel) against P2X 1 receptor, P2X 4 receptor, P2X 7 receptor, P2Y 1 receptor, or P2Y 11 receptor in TBST for 2 hours at room temperature, washed with TBST, and incubated with secondary anti-rabbit IgG (Amersham Pharmacia Biotech) in TBST for 1 hour.

Techniques: Expressing

Journal: BMC Immunology

Article Title: Expression of P2 receptors in human B cells and Epstein-Barr virus-transformed lymphoblastoid cell lines

doi: 10.1186/1471-2172-7-22

Figure Lengend Snippet: Relative expression of P2 receptors in LCLs . The relative expression of the P2X receptor gene (upper) and P2Y receptor gene (lower) in EBV-infected B cells (2 wks), LCLs, are presented (n = 4). The expression was normalized to GAPDH. P2X receptors were calibrated by P2X 1 and P2Y receptors were calibrated by P2Y 1 . Data are the mean ± SEM.

Article Snippet: The membrane was incubated with rabbit polyclonal antibodies (Alomone Labs, Jerusalem, Israel) against P2X 1 receptor, P2X 4 receptor, P2X 7 receptor, P2Y 1 receptor, or P2Y 11 receptor in TBST for 2 hours at room temperature, washed with TBST, and incubated with secondary anti-rabbit IgG (Amersham Pharmacia Biotech) in TBST for 1 hour.

Techniques: Expressing, Infection

Journal: BMC Immunology

Article Title: Expression of P2 receptors in human B cells and Epstein-Barr virus-transformed lymphoblastoid cell lines

doi: 10.1186/1471-2172-7-22

Figure Lengend Snippet: P2 receptors (P2X 1 , P2X 4 , P2X 7 , P2Y 1 , and P2Y 11 ) compared by Western blotting in B cells and LCLs . Proteins extracted from B cells and LCLs were probed with rabbit polyclonal antibodies directed against P2X 1 (60-kDa), P2X 4 (65-kDa), P2X 7 (68-kDa), P2Y 1 (66-kDa), and P2Y 11 (50-kDa) (left). Membranes were re-probed by GAPDH (40-kDa) (right). Data are representatives of 4 independent experiments.

Article Snippet: The membrane was incubated with rabbit polyclonal antibodies (Alomone Labs, Jerusalem, Israel) against P2X 1 receptor, P2X 4 receptor, P2X 7 receptor, P2Y 1 receptor, or P2Y 11 receptor in TBST for 2 hours at room temperature, washed with TBST, and incubated with secondary anti-rabbit IgG (Amersham Pharmacia Biotech) in TBST for 1 hour.

Techniques: Western Blot

Journal: BMC Immunology

Article Title: Expression of P2 receptors in human B cells and Epstein-Barr virus-transformed lymphoblastoid cell lines

doi: 10.1186/1471-2172-7-22

Figure Lengend Snippet: Sequence details for all P2X and P2Y receptor subtypes and reference (GAPDH) primers.

Article Snippet: The membrane was incubated with rabbit polyclonal antibodies (Alomone Labs, Jerusalem, Israel) against P2X 1 receptor, P2X 4 receptor, P2X 7 receptor, P2Y 1 receptor, or P2Y 11 receptor in TBST for 2 hours at room temperature, washed with TBST, and incubated with secondary anti-rabbit IgG (Amersham Pharmacia Biotech) in TBST for 1 hour.

Techniques: Sequencing

Journal: Frontiers in Physiology

Article Title: Myosin 5a in the Urinary Bladder: Localization, Splice Variant Expression, and Functional Role in Neurotransmission

doi: 10.3389/fphys.2022.890102

Figure Lengend Snippet: Purinergic responses of WT and DBA bladders. (A) Comparison of the contractile response of WT (black bar) and DBA (gray bar) bladder smooth muscle to the purinergic receptor agonist, αβmeATP (10 µM). Contractions of WT and DBA bladders were not significantly different under this condition ( N = 11 WT, N = 13 DBA; p = 0.21). (B) Western blot comparing expression of the P2X 1 purinergic receptor in WT and DBA extracts from bladder tissue devoid of mucosa. β-actin, shown in bottom panel, served as loading control. Molecular weights of mass standards (in kDa) are indicated at tick marks. (C) Quantitative data for WT (black circles) and DBA (gray squares) are graphed at right. The intensity of P2X 1 receptor immunoreactivity, normalized by intensity of β-actin, was not different between groups ( N = 3, p = 0.2).

Article Snippet: The P2X 1 purinergic receptor (P2X 1 R) antibody (APR-001, RRID:AB_2040052) was from Alomone Labs (Jerusalem, Israel).

Techniques: Western Blot, Expressing

Journal: Scientific Reports

Article Title: Indicators of the molecular pathogenesis of virulent Newcastle disease virus in chickens revealed by transcriptomic profiling of spleen

doi: 10.1038/s41598-021-96929-w

Figure Lengend Snippet: Comparison of DEGs response to NDV in the present study and other in vivo NDV infection studies. LFC stands for log 2 fold change.

Article Snippet: P2RX1 , Purinergic receptor P2X 1 , − 5.62 , Consistent with spleen of Hy-line brown at 2 dpi and lung of fayoumi at 10 dpi .

Techniques: In Vivo, Infection, Binding Assay, Activation Assay