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: 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: 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: Function

Article Title: Deletion of Mechanosensory β1-integrin From Bladder Smooth Muscle Results in Voiding Dysfunction and Tissue Remodeling

doi: 10.1093/function/zqac042

Figure Lengend Snippet: Western blotting on primary neurotransmitter receptors in BSM. (A) Muscarinic acetylcholine receptor 2 (M2), (B) muscarinic acetylcholine receptor 3 (M3), and (C) purinergic receptor P 2 X1. Upper blots show the respective antibody binding to M3, M2, and P2X 1 , while the lower blots show beta-actin staining. Quantitation by densitometry is shown beneath with receptor signals normalized to actin. Data are mean ± SD (D) qPCR data on all three receptors showing delta Ct values for mRNA extracted from 4 mice/group. Bars are mean and max–min range. * P < 0.05, ** P < 0.01 by unpaired 2-tailed t -test.

Article Snippet: Western blotting : Rabbit antibodies to β1-integrin (ab52971), and muscarinic receptor 3 (M3, ab126168) were from Abcam (Waltham, MA, USA); α1-integrin (#71 747), β3-integrin (#13 166), and β-actin (#4697) were from Cell Signaling Technology (Danvers, MA, USA); M2 (G206) was from Assay Biotech (Fremont, CA, USA) and P2X 1 (APR001) was from Alomone Laboratories (Jerusalem, Israel).

Techniques: Western Blot, Binding Assay, Staining, Quantitation Assay