Journal: Clinical Science

Article Title: Antiepileptic effects of long-term intracerebroventricular infusion of angiotensin-(1-7) in an animal model of temporal lobe epilepsy

doi: 10.1042/cs20200514

Figure Lengend Snippet: Figure 7. Effects of TLE and Ang-(1-7) treatment on hippocampal protein levels Representative Western blot bands (A) and protein levels of ACE2 (B), NEP (C), AT2R (D), AT1R (*P=0.0406 CT vs EP) (E), Mas [*P=0.0406 CT vs EP + Ang-(1-7)] (F), IL-6 (G), SOD (H), CAT (*P=0.0389 CT vs EP) (I), Bcl-2 (*P=0.0468 CT vs EP) (J), and

Article Snippet: Published by Portland Press Limited on behalf of the Biochemical Society 2265 D ow nloaded from http://portlandpress.com /clinsci/article-pdf/134/17/2263/892258/cs-2020-0514.pdf by U niversidade Federal de Sao Paulo (U N IFESP) user on 14 M ay 2024 Table 1 Host, specificity, dilution, catalog number and source of the antibodies used in the present study Antibody Host Specificity Dilution Catalog number Source ACE2 Rabbit Monoclonal 1:1000 GTX01160 GeneTex, CA, U.S.A. NEP Mouse Monoclonal 1:1000 sc-46656 Santa Cruz Biotechnology, CA, U.S.A. AT1 Mouse Monoclonal 1:500 sc-515884 Santa Cruz Biotechnology, CA, U.S.A. AT2 Rabbit Monoclonal 1:1000 M00432 Boster Biological Technology, CA, U.S.A. Mas Rabbit Polyclonal 1:500 ab66030 Abcam, MA, U.S.A. IL-6 Mouse Monoclonal 1:500 IM-0407 Imuny Biotechnology, SP, Brazil SOD Mouse Monoclonal 1:1000 sc-17767 Santa Cruz Biotechnology, CA, U.S.A. CAT Mouse Monoclonal 1:1000 LS-B2554 Lifespan Biosciences, WA, U.S.A. Bcl-2 Mouse Monoclonal 1:1000 sc-7382 Santa Cruz Biotechnology, CA, U.S.A. mTOR Mouse Monoclonal 1:1000 #4517 Cell Signaling Technology, MA, U.S.A. Phospho-mTOR Rabbit Polyclonal 1:1000 #2971 Cell Signaling Technology, MA, U.S.A. GAPDH Mouse Monoclonal 1:2000 sc-365062 Santa Cruz Biotechnology, CA, U.S.A. 2266 © 2020 The Author(s).

Techniques: Western Blot

Journal: Frontiers in Immunology

Article Title: Defining the Innate Immune Responses for SARS-CoV-2-Human Macrophage Interactions

doi: 10.3389/fimmu.2021.741502

Figure Lengend Snippet: SARS-CoV-2 restrictive infection of human MDMs. (A) Expression of SARS-CoV-2 cell entry receptor ACE2 and phenotypic surface markers CD14 and CD16, during differentiation of monocytes into macrophages, was analyzed by flow cytometry in absence or presence of captopril. (B) SARS-CoV-2 presence in MDMs. SARS-CoV-2 (MOI=0.01) was used to infect MDMs (with or without captopril). The number of virus genome equivalents per ml was measured in cell lysates and culture supernatants by RT-qPCR. Vero.STAT1 KO cells served as a positive control. (C) Transmission electron micrographs of SARS-CoV-2-challenged MDMs. ( C .i) A viral particle from a pool of SARS-CoV-2 used for the virus challenges. SARS-CoV-2-challenged MDMs [( C. ii) day 5, and ( C. iii) day 14 after viral exposure]. Red arrows, circles, and boxes demonstrate clusters of viral particles within the virus-challenged macrophages. All experiments were done at least twice with representative images depicted here. (A, B) Data are represented as mean ± SEM (n=3-6 donors). Statistical significance between groups was determined using one-way ANOVA, and p < 0.05 was considered significant (*significantly different from day 0 w/o captopril, a: significantly different from day 1 w/o captopril, b: significantly different from day 5 w/o captopril, a w : significantly different from day 1 with captopril, b w : significantly different from day 3 with captopril, c w : significantly different from day 5 with captopril). w/o: without. Scale bars: 50 nm (i and left panel of iii), 100 nm (left panel of ii), and 400 nm (right panels of ii and iii).

Article Snippet: On days 0, 1, 3, 5, and 7 during differentiation, monocytes-macrophages were stained with fluorescently-conjugated antibodies to detect human ACE2 (APC, LSBio, LS−C275129, polyclonal), CD14 (Alexa Fluor 488, eBioscience, clone 61D3), and CD16 (PE, eBioscience, eBioCB16, clone CB16), and with isotype-matched antibodies serving as negative controls.

Techniques: Infection, Expressing, Flow Cytometry, Quantitative RT-PCR, Positive Control, Transmission Assay

Journal: European Journal of Clinical Investigation

Article Title: Angiotensin‐converting enzyme 2 and transmembrane protease serine 2 in female and male patients with end‐stage kidney disease

doi: 10.1111/eci.13786

Figure Lengend Snippet: Serum concentration of soluble angiotensin‐converting enzyme 2 (ACE2) in (A) controls versus patients with end‐stage kidney disease (ESKD), (B, C) sex divided differences into controls and ESKD subjects. Results are expressed as the median and interquartile range (IQR). Significance p < 0.05

Article Snippet: The diluted primary antibody ACE2 (1:200; Nordic BioSite, cat no: CSB‐PA866317LA01HU‐50, Sweden), TMPRSS2 (1:400; cat no HPA035787, Sigma Aldrich) or CD31 (1:100, cat no: 553370, BD Biosciences) with PBS and 5% goat serum was added on the slide and incubated overnight in 4°C.

Techniques: Concentration Assay

Journal: European Journal of Clinical Investigation

Article Title: Angiotensin‐converting enzyme 2 and transmembrane protease serine 2 in female and male patients with end‐stage kidney disease

doi: 10.1111/eci.13786

Figure Lengend Snippet: Correlation of soluble ACE2 with clinical and other biochemical parameters in controls and ESKD patients

Article Snippet: The diluted primary antibody ACE2 (1:200; Nordic BioSite, cat no: CSB‐PA866317LA01HU‐50, Sweden), TMPRSS2 (1:400; cat no HPA035787, Sigma Aldrich) or CD31 (1:100, cat no: 553370, BD Biosciences) with PBS and 5% goat serum was added on the slide and incubated overnight in 4°C.

Techniques: Control

Journal: European Journal of Clinical Investigation

Article Title: Angiotensin‐converting enzyme 2 and transmembrane protease serine 2 in female and male patients with end‐stage kidney disease

doi: 10.1111/eci.13786

Figure Lengend Snippet: Soluble ACE2 multivariable linear regression analysis in ESKD participants

Article Snippet: The diluted primary antibody ACE2 (1:200; Nordic BioSite, cat no: CSB‐PA866317LA01HU‐50, Sweden), TMPRSS2 (1:400; cat no HPA035787, Sigma Aldrich) or CD31 (1:100, cat no: 553370, BD Biosciences) with PBS and 5% goat serum was added on the slide and incubated overnight in 4°C.

Techniques:

Journal: European Journal of Clinical Investigation

Article Title: Angiotensin‐converting enzyme 2 and transmembrane protease serine 2 in female and male patients with end‐stage kidney disease

doi: 10.1111/eci.13786

Figure Lengend Snippet: Soluble ACE2 multivariable linear regression analysis in non‐CKD control participants

Article Snippet: The diluted primary antibody ACE2 (1:200; Nordic BioSite, cat no: CSB‐PA866317LA01HU‐50, Sweden), TMPRSS2 (1:400; cat no HPA035787, Sigma Aldrich) or CD31 (1:100, cat no: 553370, BD Biosciences) with PBS and 5% goat serum was added on the slide and incubated overnight in 4°C.

Techniques: Control

Journal: European Journal of Clinical Investigation

Article Title: Angiotensin‐converting enzyme 2 and transmembrane protease serine 2 in female and male patients with end‐stage kidney disease

doi: 10.1111/eci.13786

Figure Lengend Snippet: Soluble ACE2 levels in patients with end‐stage kidney disease (ESKD) according to treatment and presence of comorbidities cardiovascular disease (CVD), diabetes mellitus (DM) and combined CVD + DM. Results are expressed as the median and interquartile range (IQR). Significance p < 0.05

Article Snippet: The diluted primary antibody ACE2 (1:200; Nordic BioSite, cat no: CSB‐PA866317LA01HU‐50, Sweden), TMPRSS2 (1:400; cat no HPA035787, Sigma Aldrich) or CD31 (1:100, cat no: 553370, BD Biosciences) with PBS and 5% goat serum was added on the slide and incubated overnight in 4°C.

Techniques:

Journal: European Journal of Clinical Investigation

Article Title: Angiotensin‐converting enzyme 2 and transmembrane protease serine 2 in female and male patients with end‐stage kidney disease

doi: 10.1111/eci.13786

Figure Lengend Snippet: Angiotensin‐converting enzyme 2 (ACE2) expression in resistance artery sections from patients with end‐stage kidney disease (ESKD) and controls. (A) Representative immunofluorescence staining of ACE2, (B) Statistical analysis of ACE2 expression between controls ( n = 15) and ESKD ( n = 23), (C) Representative immunofluorescence staining of ACE2 expression and CD31 endothelial cell marker expression in resistance artery from patients with ESKD. ACE2 (red), CD31 (green) and nuclear DAPI staining (blue). Co‐localization of ACE2 and CD31 expression is seen in yellow. Bar = 100 µm. Significance p < 0.05; ESKD, end‐stage kidney disease

Article Snippet: The diluted primary antibody ACE2 (1:200; Nordic BioSite, cat no: CSB‐PA866317LA01HU‐50, Sweden), TMPRSS2 (1:400; cat no HPA035787, Sigma Aldrich) or CD31 (1:100, cat no: 553370, BD Biosciences) with PBS and 5% goat serum was added on the slide and incubated overnight in 4°C.

Techniques: Expressing, Immunofluorescence, Staining, Marker

Journal: European Journal of Clinical Investigation

Article Title: Angiotensin‐converting enzyme 2 and transmembrane protease serine 2 in female and male patients with end‐stage kidney disease

doi: 10.1111/eci.13786

Figure Lengend Snippet: Sex‐divided analysis of angiotensin‐converting enzyme 2 (ACE2) expression in resistance artery sections from patients with end‐stage kidney disease (ESKD) and controls. (A) Representative immunofluorescence staining of ACE2 (red) in male and female controls, Bar = 100µm, (B) Sex divided statistical analysis of ACE2 expression in male vs female controls ( n = 6/9, respectively), and male versus female ESKD patients ( n = 13/10, respectively). M, male; F, female; Significance p < 0.05; ESKD, end‐stage kidney disease

Article Snippet: The diluted primary antibody ACE2 (1:200; Nordic BioSite, cat no: CSB‐PA866317LA01HU‐50, Sweden), TMPRSS2 (1:400; cat no HPA035787, Sigma Aldrich) or CD31 (1:100, cat no: 553370, BD Biosciences) with PBS and 5% goat serum was added on the slide and incubated overnight in 4°C.

Techniques: Expressing, Immunofluorescence, Staining

Journal: European Journal of Clinical Investigation

Article Title: Angiotensin‐converting enzyme 2 and transmembrane protease serine 2 in female and male patients with end‐stage kidney disease

doi: 10.1111/eci.13786

Figure Lengend Snippet: Angiotensin‐converting enzyme 2 (ACE2) expression in resistance artery sections from patients with end‐stage kidney disease (ESKD) with or without ACE‐inhibitor/angiotensin receptor blocker (ARB) treatment. (A) Representative immunofluorescence staining of ACE2 (red) in ACE‐inhibitor/ARB treated and non‐treated arteries from ESKD patients, Bar = 100µm, (B) Statistical analysis of ACE2 expression in ACE‐inhibitor/ARB treated ( n = 12) and non‐treated ( n = 11) ESKD; Significance p < 0.05

Article Snippet: The diluted primary antibody ACE2 (1:200; Nordic BioSite, cat no: CSB‐PA866317LA01HU‐50, Sweden), TMPRSS2 (1:400; cat no HPA035787, Sigma Aldrich) or CD31 (1:100, cat no: 553370, BD Biosciences) with PBS and 5% goat serum was added on the slide and incubated overnight in 4°C.

Techniques: Expressing, Immunofluorescence, Staining

Journal: European Journal of Clinical Investigation

Article Title: Angiotensin‐converting enzyme 2 and transmembrane protease serine 2 in female and male patients with end‐stage kidney disease

doi: 10.1111/eci.13786

Figure Lengend Snippet: Transmembrane protease serine 2 (TMPRSS2) expression in resistance artery sections (A + B), and adipose tissue (C + D) from patients with end‐stage kidney disease (ESKD) and controls. (A ) Representative immunofluorescence staining of TMPRSS2 in arteries, (B) Statistical analysis of TMPRSS2 expression between controls ( n = 15) and ESKD ( n = 23) in resistance arteries, (C) Representative immunofluorescence staining of TMPRSS2 in adipose tissue, (D) Statistical analysis of TMPRSS2 expression between control ( n = 12) and ESKD ( n = 11) in adipose tissue. ACE2 (red) and nuclear staining with DAPI (blue). Bar = 100 µm. Significance p < 0.05; ESKD, end‐stage kidney disease

Article Snippet: The diluted primary antibody ACE2 (1:200; Nordic BioSite, cat no: CSB‐PA866317LA01HU‐50, Sweden), TMPRSS2 (1:400; cat no HPA035787, Sigma Aldrich) or CD31 (1:100, cat no: 553370, BD Biosciences) with PBS and 5% goat serum was added on the slide and incubated overnight in 4°C.

Techniques: Expressing, Immunofluorescence, Staining, Control

Journal: International Journal of Molecular Medicine

Article Title: High glucose activates the alternative ACE2/Ang-(1-7)/Mas and APN/Ang IV/IRAP RAS axes in pancreatic β-cells

doi: 10.3892/ijmm.2013.1469

Figure Lengend Snippet: Schematic illustration of RAS consisting of the classical Ang II/ACE/AT1R axis and AT2R, as well as the two alternative axes, ACE2/Ang-( 1 - 7 )/Mas and APN/Ang IV/IRAP. RAS, renin angiotensin system; AGT, angiotensinogen; Ang, angiotensin; AT1bR, angiotensin II type 1b receptor; AT2R, angiotensin II type 2 receptor; ACE, angiotensin-converting enzyme; NEP, neutral endopeptidase 24.11; PEP, prolyl endopeptidase; Mas, Mas receptor; APA/APN, aminopeptidases A and N; IRAP, insulin-regulated aminopeptidase.

Article Snippet: The antibodies used, antibody dilutions and dilution buffers for western blotting were as follows: rabbit anti ACE2 (LS-B439; Biozol, Eching, Germany) 1:10,000, mouse anti-NEP (CD10; ab951) 1:1,000, goat anti-APA (BP1; ab36122) 1:2,000, rabbit anti-APN (CD13; ab108310; all from Abcam Cambridge, UK) 1:2,000, rabbit anti-IRAP (#6918; NEB; Cell Signaling Technology, Frankfurt/Main, Germany) 1:2,000, all in TBST/5% BSA/0.03% NaN 3 ; rabbit anti-Mas (LS-B2564; Biozol) 1:2,000 in PBS/5% skimmed milk/0.03% NaN 3 ; goat anti -actin (I-19, SC-1616; Santa Cruz Biotechnology, Inc., Heidelberg, Germany) 1:500, rabbit anti-RPLP0 (Biozol) 1:2,000, both in TBST/5% BSA/0.03% NaN 3 .

Techniques:

Journal: International Journal of Molecular Medicine

Article Title: High glucose activates the alternative ACE2/Ang-(1-7)/Mas and APN/Ang IV/IRAP RAS axes in pancreatic β-cells

doi: 10.3892/ijmm.2013.1469

Figure Lengend Snippet: Effects of glucose on the mRNA expression of renin-angiotensin system (RAS) components. Elevated concentrations of glucose did not alter the mRNA levels of (A) angiotensin-converting enzyme (ACE), (B) angiotensin II type 1b receptor (AT1bR), or (C) angiotensin II type 2 receptor (AT2R) mRNA levels in BRIN-BD11 cells after 24 h, whereas the levels of (D) ACE2, (E) neutral endopeptidase 24.11 (NEP), (F) Mas, (G) aminopeptidase A (APA), (H) aminopeptidase N (APN) and (I) insulin-regulated aminopeptidase (IRAP) were dose-dependently increased. mRNA levels were determined by quantitative PCR. Data were evaluated and normalized to Rpl13a mRNA expression using the ΔΔ Cq method. Data are presented as box plots with medians, quartiles and an interquartile range (IQR) ± 1.5 × IQR of 5 independent experiments [Mann-Whitney * P<0.05, ** P<0.01, vs. control (Co.)].

Article Snippet: The antibodies used, antibody dilutions and dilution buffers for western blotting were as follows: rabbit anti ACE2 (LS-B439; Biozol, Eching, Germany) 1:10,000, mouse anti-NEP (CD10; ab951) 1:1,000, goat anti-APA (BP1; ab36122) 1:2,000, rabbit anti-APN (CD13; ab108310; all from Abcam Cambridge, UK) 1:2,000, rabbit anti-IRAP (#6918; NEB; Cell Signaling Technology, Frankfurt/Main, Germany) 1:2,000, all in TBST/5% BSA/0.03% NaN 3 ; rabbit anti-Mas (LS-B2564; Biozol) 1:2,000 in PBS/5% skimmed milk/0.03% NaN 3 ; goat anti -actin (I-19, SC-1616; Santa Cruz Biotechnology, Inc., Heidelberg, Germany) 1:500, rabbit anti-RPLP0 (Biozol) 1:2,000, both in TBST/5% BSA/0.03% NaN 3 .

Techniques: Expressing, Real-time Polymerase Chain Reaction, MANN-WHITNEY, Control

Journal: International Journal of Molecular Medicine

Article Title: High glucose activates the alternative ACE2/Ang-(1-7)/Mas and APN/Ang IV/IRAP RAS axes in pancreatic β-cells

doi: 10.3892/ijmm.2013.1469

Figure Lengend Snippet: Effects of glucose on enzymatic activities of renin-angiotensin system (RAS) proteases. Exposure of BRIN-BD11 cells to elevated concentrations of glucose led to increased enzymatic activities of (A) angiotensin-converting enzyme (ACE)2, (B) aminopeptidase A (APA), (C) aminopeptidase N (APN) and (D) insulin-regulated aminopeptidase (IRAP) after 24 h. Protease activities were determined by the incubation of viable cells with chromogenic or fluorogenic (ACE2) substrates. The activities were calculated per one million cells. Illustrated are boxplots with medians, quartiles and an interquartile range (IQR) ± 1.5 × IQR of 4 independent experiments [Mann-Whitney ** P<0.01, * P<0.05, vs. control (Co.)].

Article Snippet: The antibodies used, antibody dilutions and dilution buffers for western blotting were as follows: rabbit anti ACE2 (LS-B439; Biozol, Eching, Germany) 1:10,000, mouse anti-NEP (CD10; ab951) 1:1,000, goat anti-APA (BP1; ab36122) 1:2,000, rabbit anti-APN (CD13; ab108310; all from Abcam Cambridge, UK) 1:2,000, rabbit anti-IRAP (#6918; NEB; Cell Signaling Technology, Frankfurt/Main, Germany) 1:2,000, all in TBST/5% BSA/0.03% NaN 3 ; rabbit anti-Mas (LS-B2564; Biozol) 1:2,000 in PBS/5% skimmed milk/0.03% NaN 3 ; goat anti -actin (I-19, SC-1616; Santa Cruz Biotechnology, Inc., Heidelberg, Germany) 1:500, rabbit anti-RPLP0 (Biozol) 1:2,000, both in TBST/5% BSA/0.03% NaN 3 .

Techniques: Incubation, MANN-WHITNEY, Control