Journal: PLoS ONE

Article Title: The Effects of Angiotensin II and Angiotensin-(1–7) in the Rostral Ventrolateral Medulla of Rats on Stress-Induced Hypertension

doi: 10.1371/journal.pone.0070976

Figure Lengend Snippet: ACE upregulation and/or ACE2 downregulation result in Ang II↑ and/or Ang-(1–7)↓. Ang, Angiotensin; ACE, angiotensin-converting enzyme; ACE2, angiotensin-converting enzyme homolog; AT 1 R, Ang II type 1 receptor; AT 2 R, Ang II type 2 receptor; SIH, stress-induced hypertension

Article Snippet: Primary antibodies were rabbit polyclonal antibodies against AT 1 R (1∶2000) (Alomone, Israel), rabbit polyclonal antibody against AT 2 R (1∶1000) (Santa Cruz, USA) , rabbit polyclonal antibodies against Mas Receptor (1∶2000) (Alomone, Israel) , mouse monoclonal antibody against ACE (1∶1000) (abcam, USA) , goat polyclonal antibody against ACE2 (1∶1000) (Santa Cruz, USA), and mouse polyclonal antibodies against β-actin (1∶3000) (as an internal standard from Beyotime Institute of Biotechnology, Haimen, China).

Techniques:

Journal: Cells

Article Title: Increased K V 2.1 Channel Clustering Underlies the Reduction of Delayed Rectifier K + Currents in Hippocampal Neurons of the Tg2576 Alzheimer’s Disease Mouse

doi: 10.3390/cells11182820

Figure Lengend Snippet: Effect of the intracellular diffusion of the anti-K V 2.1 monoclonal antibody on I DR recorded in the WT and Tg2576 primary hippocampal neurons. ( A ) Time-dependent inhibitory effect of the anti-K V 2.1 monoclonal antibody (10 µg mL − 1 ) on I DR recorded in WT and Tg2576 primary hippocampal pyramidal neurons. ( B ) Quantification of I DR densities, at +40 mV, before (Control) and after 6 min of the anti-K V 2.1 intracellular diffusion (+K V 2.1Ab). Data are expressed as percentage of WT and Tg2576 internal controls. Values are expressed as mean ± SEM of 3 independent experimental sessions. * p < 0.001 vs. internal controls. ( C ) Superimposed representative traces of I DR at +40 mV recorded before and after 6 min of intracellular diffusion of the anti-K V 2.1 antibody ( top ), and representative traces resulting from the subtraction of the currents resistant to the anti-K V 2.1 antibody from the initial currents ( bottom ) in WT ( left ) and Tg2576 ( right ) neurons. ( D ) Representative traces of I DR recorded in control conditions and upon 6 min of intracellular diffusion of the anti-K V 2.1 monoclonal antibody (10 µg mL − 1 ) in WT ( left ) and Tg2576 ( right ) primary hippocampal pyramidal neurons. Protocols are shown in the lower part of the panel. ( E ) I/V relationships for the I DR recorded in control conditions and upon 6 min of intracellular diffusion of the anti-K V 2.1 antibody in the WT ( left ) and Tg2576 primary hippocampal neurons ( right ). ( F ) Steady-state activation curves of I DR recorded in WT and Tg2576 hippocampal neurons in control conditions and in WT neurons upon 6 min of intracellular diffusion of the anti-K V 2.1 antibody. Values are expressed as mean ± SEM of 3 independent experimental sessions. * p < 0.001 vs. WT.

Article Snippet: To this aim, both the Tg2576 and WT hippocampal neurons were fixed and stained with the anti-K V 2.1 rabbit polyclonal antibody (1:1000, Alomone Labs, Jerusalem, Israel) and subjected to imaging on a confocal microscope.

Techniques: Diffusion-based Assay, Activation Assay

Journal: Cells

Article Title: Increased K V 2.1 Channel Clustering Underlies the Reduction of Delayed Rectifier K + Currents in Hippocampal Neurons of the Tg2576 Alzheimer’s Disease Mouse

doi: 10.3390/cells11182820

Figure Lengend Snippet: K V 2.1 clustering and effect of glutamate on I DR in WT and Tg2576 primary hippocampal pyramidal neurons. ( A ) Representative confocal images of pyramidal neurons from Tg2576 ( c , d ) and WT ( a , b ) primary hippocampal cultures stained with anti-K V 2.1 antibody (green). On the right, a magnification of neuronal somata from the confocal images shown on the left. ( B ) Time-dependent stimulatory effect of glutamate (10 µM) on I DR recorded in WT and Tg2576 primary hippocampal pyramidal neurons. ( C ) Quantification of I DR densities, at +40 mV, before (Control) and after 11 min of 10 µM glutamate exposure (Glu). Data are represented as percentage of WT and Tg2576 internal controls. Values are expressed as mean ± SEM of 3 independent experimental sessions. * p < 0.001 vs. internal controls. ( D ) Representative traces of I DR recorded in control conditions and upon 11 min of 10 µM glutamate exposure in WT ( left ) and Tg2576 ( right ) primary hippocampal pyramidal neurons. Protocols are shown in the lower part of the panel. ( E ) I/V relationships for the I DR recorded in control conditions and upon 11 min of 10 µM glutamate exposure in the WT ( left ) and Tg2576 primary hippocampal neurons ( right ). ( F ) Steady-state activation curves of I DR recorded in WT and Tg2576 hippocampal neurons in control conditions and in Tg2576 neurons upon 11 min of 10 µM glutamate exposure. Values are expressed as mean ± SEM of 3 independent experimental sessions.

Article Snippet: To this aim, both the Tg2576 and WT hippocampal neurons were fixed and stained with the anti-K V 2.1 rabbit polyclonal antibody (1:1000, Alomone Labs, Jerusalem, Israel) and subjected to imaging on a confocal microscope.

Techniques: Staining, Activation Assay

Journal: Cells

Article Title: Increased K V 2.1 Channel Clustering Underlies the Reduction of Delayed Rectifier K + Currents in Hippocampal Neurons of the Tg2576 Alzheimer’s Disease Mouse

doi: 10.3390/cells11182820

Figure Lengend Snippet: Effect of the anti-K V 2.1 monoclonal antibody on the positive modulation of I DR by glutamate in WT and Tg2576 primary hippocampal neurons. ( A ) Superimposed representative traces of I DR at +40 mV recorded before (Control) and after 11 min of 10 µM glutamate (Glu), and representative traces recorded before and after the intracellular diffusion of the anti-K V 2.1 antibody and after 11 min of 10 µM glutamate exposure in the presence of the anti-K V 2.1 antibody. ( B ) Quantification of I DR densities, at +40 mV, in control conditions and upon 11 min of 10 µM glutamate exposure with (+K V 2.1Ab) and without the anti-K V 2.1 antibody (−K V 2.1Ab) in the recording pipette. Data are expressed as percentage of WT and Tg2576 internal controls. Values are expressed as mean ± SEM of 3 independent experimental sessions. * p < 0.001 vs. control; ** p < 0.001 vs. GLU + K V 2.1Ab.

Article Snippet: To this aim, both the Tg2576 and WT hippocampal neurons were fixed and stained with the anti-K V 2.1 rabbit polyclonal antibody (1:1000, Alomone Labs, Jerusalem, Israel) and subjected to imaging on a confocal microscope.

Techniques: Diffusion-based Assay, Transferring

Journal: Biomedicines

Article Title: Vascular Effects of Low-Dose ACE2 Inhibitor MLN-4760—Benefit or Detriment in Essential Hypertension?

doi: 10.3390/biomedicines10010038

Figure Lengend Snippet: The equilibrium angiotensin peptides levels in plasma.

Article Snippet: Membranes were blocked with 5% nonfat milk for 1 h at room temperature in Tris-buffer solution (pH 7.6) containing 0.1% Tween-20 and probed against the following primary antibodies: rabbit polyclonal anti-endothelial NOS, anti-neuronal NOS and anti-β-actin (Abcam, Cambridge, UK); rabbit polyclonal anti-inducible NOS (Bio-Rad, Inc., Hercules, CA, USA) rabbit polyclonal anti-angiotensin(1–7) Mas receptor (Alomone Labs, Jerusalem, Israel); rabbit polyclonal CBS and mouse monoclonal anti-CSE antibodies (Proteintech, Manchester, UK); and rabbit monoclonal anti-ACE2 (Invitrogen, Waltham, MA, USA) overnight at 4 °C.

Techniques:

Journal: Biomedicines

Article Title: Vascular Effects of Low-Dose ACE2 Inhibitor MLN-4760—Benefit or Detriment in Essential Hypertension?

doi: 10.3390/biomedicines10010038

Figure Lengend Snippet: The correlation between the plasma level of the total alternative renin angiotensin system activity (Alt-S) and the weight of the visceral fat ( a ) and body weight ( b ). The correlation between the plasma level of angiotensin 1–7 (Ang 1–7) and the weight of the visceral fat ( c ) and body weight ( d ) in SHRs and SHRs treated with MLN-4760 (SHR + MLN).

Article Snippet: Membranes were blocked with 5% nonfat milk for 1 h at room temperature in Tris-buffer solution (pH 7.6) containing 0.1% Tween-20 and probed against the following primary antibodies: rabbit polyclonal anti-endothelial NOS, anti-neuronal NOS and anti-β-actin (Abcam, Cambridge, UK); rabbit polyclonal anti-inducible NOS (Bio-Rad, Inc., Hercules, CA, USA) rabbit polyclonal anti-angiotensin(1–7) Mas receptor (Alomone Labs, Jerusalem, Israel); rabbit polyclonal CBS and mouse monoclonal anti-CSE antibodies (Proteintech, Manchester, UK); and rabbit monoclonal anti-ACE2 (Invitrogen, Waltham, MA, USA) overnight at 4 °C.

Techniques: Activity Assay

Journal: Peptides

Article Title: Impact of angiotensin II type 1 and G-protein-coupled Mas receptor expression on the pulmonary performance of patients with idiopathic pulmonary fibrosis

doi: 10.1016/j.peptides.2020.170384

Figure Lengend Snippet: Protein expression of AT1 and Mas receptors in lung tissue (n = 12). A: Expression of AT1 and Mas receptors in idiopathic pulmonary fibrosis lung tissue and control evaluated by western blot. B: Protein quantification of AT1 receptor in idiopathic pulmonary fibrosis and control lung tissue. C: Protein quantification of Mas receptor in idiopathic pulmonary fibrosis and control lung tissue.

Article Snippet: Afterwards the membrane was stained with a 1:500 concentration of rabbit polyclonal anti-human anti-angiotensin II type-1 receptor antibody/AGTR1 (AAR-011, Alomone®, Israel) and a 1:250 concentration of rabbit polyclonal anti-human anti-angiotensin-(1–7) Mas receptor antibody (AAR-013, Alomone®, Israel) or mouse anti-human β-actin monoclonal antibody (A2228, Sigma Aldrich®, Germany), followed by secondary staining with a 1:1000 concentration of rabbit anti-mouse IgG (H + L)-HRP antibody (ThermoFisher Scientific®, MA, USA).

Techniques: Expressing, Western Blot

Journal: Peptides

Article Title: Impact of angiotensin II type 1 and G-protein-coupled Mas receptor expression on the pulmonary performance of patients with idiopathic pulmonary fibrosis

doi: 10.1016/j.peptides.2020.170384

Figure Lengend Snippet: Scatter plots of correlation analysis between spirometry values and AT1 receptor quantification (n = 12). FEV 1 =forced expiratory volume in the first second; FVC = forced vital capacity.

Article Snippet: Afterwards the membrane was stained with a 1:500 concentration of rabbit polyclonal anti-human anti-angiotensin II type-1 receptor antibody/AGTR1 (AAR-011, Alomone®, Israel) and a 1:250 concentration of rabbit polyclonal anti-human anti-angiotensin-(1–7) Mas receptor antibody (AAR-013, Alomone®, Israel) or mouse anti-human β-actin monoclonal antibody (A2228, Sigma Aldrich®, Germany), followed by secondary staining with a 1:1000 concentration of rabbit anti-mouse IgG (H + L)-HRP antibody (ThermoFisher Scientific®, MA, USA).

Techniques: