Journal: Hypertension

Article Title: Identification of Bona Fide Alternative Renin Transcripts Expressed Along Cortical Tubules and Potential Roles in Promoting Insulin Resistance In Vivo Without Significant Plasma Renin Activity Elevation

doi: 10.1161/hypertensionaha.114.03394

Figure Lengend Snippet: Figure 7. A, The results of quantitative reverse transcription polymerase chain reaction analyses of inflammatory molecules in heart, kidney, and liver of both ARen2TG and wild-type littermates are shown as relative abundance, using GAPDH expression as an internal control. *, **, #, ##Significantly elevated compared with the expressions of wild-type littermates. Repeated measure ANOVA and subsequent post hoc analyses were performed. B, Representative results of in situ histological examinations for livers of both transgenic mice and wild-type littermate using rabbit polyclonal anti-angiotensin II antibodies (NBP1-30027; Novus Biologicals, LLC, Littleton, CO) were shown. a–c, Enhanced immunohistochemical staining in livers of transgenic mice compared with those of wild-type littermates (Bd), suggesting enhanced angiotensin II production in the organs.

Article Snippet: We performed experiments to compare ARen2TG mice with wild-type littermates in terms of following phenotypes: measurement of blood pressure,16,17 measurement of plasma renin activity,16,18,19 the results of in vivo near-infrared imaging using the renin substrate analog ReninSense 680 FAST (Perkin Elmer, Waltham, MA),20 the results of insulin tolerance tests,21 fasting plasma glucose levels, the expressions of inflammatory molecules in various tissues on quantitative RT-PCR analyses,16,18,19 and in situ histological examinations for livers of both transgenic mice and wild-type littermate using rabbit polyclonal anti-angiotensin II antibodies (NBP1-30027; Novus Biologicals, LLC, Littleton, CO).22,23 All experiments were performed according to the protocols of manufacturer, the references, and our reports as specified above.

Techniques: Reverse Transcription, Polymerase Chain Reaction, Expressing, Control, In Situ, Transgenic Assay, Immunohistochemical staining, Staining

Journal: Pflugers Archiv

Article Title: Renal angiotensin I-converting enzyme-deficient mice are protected against aristolochic acid nephropathy

doi: 10.1007/s00424-022-02779-4

Figure Lengend Snippet: a Renal angiotensinogen (AGT) protein abundance, b renal renin protein abundance normalized to total protein, and c Western blot from kidney tissue using an antibody against AGT and d against renin in the wildtype and ACE −/− mice treated with vehicle or aristolochic acid I (AAI). ACE −/− , C57BL/6J- tm (ACE3/3) mice, n = 6–10 per group, two-way ANOVA: a treatment p < 0.001; strain n.s. ; interaction n.s. ; b treatment p < 0.001; strain p < 0.001; interaction p < 0.05; Bonferroni test: *** p < 0.0001 vs. vehicle; ## p < 0.01 vs. wildtype

Article Snippet: After separation, proteins were transferred to a nitrocellulose membrane, blocked with 5% bovine serum albumin or Roti Block (Roth, Karlsruhe, Germany), and incubated with antibodies against AGT (1:4000, R and D Systems Cat# AF6966, RRID:AB_10971952, Inc., Minneapolis, USA), renin (1:2000, ABIN301824, antibodies-online, Aachen, Germany), ACE (1:1000, orb216086; Biorbyt Ltd., Cambridge, UK), ACE2 (1:1000, Abnova Cat# PAB13443, RRID:AB_10549736, Taipei City, Taiwan) and Mas (1:2000, Novus Cat# NBP1-78,444, RRID:AB_11039164, Wiesbaden-Nordenstadt, Germany), respectively, followed by a secondary horseradish peroxidase (HRP)-conjugated antibody (goat anti-rabbit HRP Conjugate, Bio-Rad Cat# 170–6515, RRID:AB_11125142, Munich, Germany or rabbit anti-goat HRP Conjugate, Millipore Cat# AP106P, RRID:AB_9241, Darmstadt, Germany).

Techniques: Quantitative Proteomics, Western Blot

Journal: Journal of Vascular Anomalies

Article Title: Expression of Angiotensin II Receptor 2 in Microcystic Lymphatic Malformation

doi: 10.1097/jova.0000000000000020

Figure Lengend Snippet: Figure 1. A representative hematoxylin and eosin–stained section (A) and immunohistochemical-stained sections (B–H) of microcystic lymphatic malformation showing characteristic ectatic lymphatic channels lined by flat endothelial cells (A) which stained positively for D2–40 (B, brown). Angiotensinogen (C, brown), renin (D, brown), PRR (E, brown), ACE (F, brown), and ACE2 (G, brown) were not detected on the lesional lymphatic vessels. AT2R (H, brown) was present on the endothelium of the lesional lymphatic vessels. Nuclei were counter-stained with hematoxylin (A–H, blue). Original magnification: ×200, inset ×400. ACE indicates angiotensin-converting enzyme; AT2R, angiotensin II receptor 2; PRR, prorenin receptor.

Article Snippet: Immunohistochemical staining was then performed with primary antibodies for angiotensinogen (1:50, cat#79299, Cell Signaling, Danvers, MA), renin (1:500, cat#14291-1-AP, Proteintech, Rosemont, IL), PRR (1:500; cat#ab40790, Abcam, Cambridge, United Kingdom), ACE (1:100; cat#MCA2056, Bio-Rad, Hercules, CA), ACE2 (1:200; cat#MAB933 R&D Systems, Minneapolis, MN), and angiotensin II receptor 2 (AT2R; 1:2000; cat#NBP1-77368, Novus Biologicals, Littleton, CO).

Techniques: Staining, Immunohistochemical staining

Journal: Journal of Vascular Anomalies

Article Title: Expression of Angiotensin II Receptor 2 in Microcystic Lymphatic Malformation

doi: 10.1097/jova.0000000000000020

Figure Lengend Snippet: Figure 3. Full-length western blot images of total protein extracted from microcystic lymphatic malformation tissue samples from 6 patients demonstrating the presence of angiotensinogen (A, red) in all 6 samples at the expected size of ~60 kDa. PRR (B, red) was detected in all samples at the appropriate size of ~35 kDa, with bands representing the soluble form of PRR at ~27 kDa in 4 samples. ACE (C, red) was detected in 5 of the 6 tissue samples at ~195 kDa. ACE2 (D, red) was detected in the positive control, but none of the samples. AT2R (E, red) was similarly confirmed in the positive control but was not detected in the tissue samples. α-tubulin loading control (F, red) confirmed approximate equal loading. ACE indicates angiotensin-converting enzyme; AT2R, angiotensin II receptor 2; PRR, prorenin receptor.

Article Snippet: Immunohistochemical staining was then performed with primary antibodies for angiotensinogen (1:50, cat#79299, Cell Signaling, Danvers, MA), renin (1:500, cat#14291-1-AP, Proteintech, Rosemont, IL), PRR (1:500; cat#ab40790, Abcam, Cambridge, United Kingdom), ACE (1:100; cat#MCA2056, Bio-Rad, Hercules, CA), ACE2 (1:200; cat#MAB933 R&D Systems, Minneapolis, MN), and angiotensin II receptor 2 (AT2R; 1:2000; cat#NBP1-77368, Novus Biologicals, Littleton, CO).

Techniques: Western Blot, Positive Control, Control

Journal: PLoS ONE

Article Title: Improved Assay for Quantifying a Redox Form of Angiotensinogen as a Biomarker for Pre-Eclampsia: A Case-Control Study

doi: 10.1371/journal.pone.0135905

Figure Lengend Snippet: Total angiotensinogen plasma level is the sum of the plasma levels of the oxidized and reduced forms of angiotensinogen.

Article Snippet: Mouse anti-human angiotensinogen monoclonal antibody (clone 369439) was purchased from R&D Systems, Minneapolis, MN.

Techniques: Clinical Proteomics

Journal: PLoS ONE

Article Title: Improved Assay for Quantifying a Redox Form of Angiotensinogen as a Biomarker for Pre-Eclampsia: A Case-Control Study

doi: 10.1371/journal.pone.0135905

Figure Lengend Snippet: Biotin-MPB is added to plasma which labels free thiol (SH) angiotensinogen (ATG). The unincorporated Biotin-MPB is removed by acetone precipitation. Biotin-MPB labelled free thiols of ATG are specifically bound by streptavidin coated plates and bound free thiol ATG is quantified with a specific monoclonal antibody to ATG.

Article Snippet: Mouse anti-human angiotensinogen monoclonal antibody (clone 369439) was purchased from R&D Systems, Minneapolis, MN.

Techniques: Clinical Proteomics

Journal: PLoS ONE

Article Title: Improved Assay for Quantifying a Redox Form of Angiotensinogen as a Biomarker for Pre-Eclampsia: A Case-Control Study

doi: 10.1371/journal.pone.0135905

Figure Lengend Snippet: The amount of total plasma angiotensinogen was derived from a standard curve constructed from known concentrations of purified angiotensinogen. NS = Not Significant.

Article Snippet: Mouse anti-human angiotensinogen monoclonal antibody (clone 369439) was purchased from R&D Systems, Minneapolis, MN.

Techniques: Clinical Proteomics, Derivative Assay, Construct, Purification

Journal: PLoS ONE

Article Title: Improved Assay for Quantifying a Redox Form of Angiotensinogen as a Biomarker for Pre-Eclampsia: A Case-Control Study

doi: 10.1371/journal.pone.0135905

Figure Lengend Snippet: Levels of free thiol angiotensinogen are expressed as a percentage of the in-house standard (**** p<0.0001).

Article Snippet: Mouse anti-human angiotensinogen monoclonal antibody (clone 369439) was purchased from R&D Systems, Minneapolis, MN.

Techniques:

Journal: PLoS ONE

Article Title: Improved Assay for Quantifying a Redox Form of Angiotensinogen as a Biomarker for Pre-Eclampsia: A Case-Control Study

doi: 10.1371/journal.pone.0135905

Figure Lengend Snippet: ROC analysis for total and free thiol angiotensinogen.

Article Snippet: Mouse anti-human angiotensinogen monoclonal antibody (clone 369439) was purchased from R&D Systems, Minneapolis, MN.

Techniques:

Journal: PLoS ONE

Article Title: Improved Assay for Quantifying a Redox Form of Angiotensinogen as a Biomarker for Pre-Eclampsia: A Case-Control Study

doi: 10.1371/journal.pone.0135905

Figure Lengend Snippet: ROC curve for total angiotensinogen.

Article Snippet: Mouse anti-human angiotensinogen monoclonal antibody (clone 369439) was purchased from R&D Systems, Minneapolis, MN.

Techniques:

Journal: PLoS ONE

Article Title: Improved Assay for Quantifying a Redox Form of Angiotensinogen as a Biomarker for Pre-Eclampsia: A Case-Control Study

doi: 10.1371/journal.pone.0135905

Figure Lengend Snippet: ROC curve for free thiol angiotensinogen.

Article Snippet: Mouse anti-human angiotensinogen monoclonal antibody (clone 369439) was purchased from R&D Systems, Minneapolis, MN.

Techniques:

Journal: Revista da Associação Médica Brasileira

Article Title: Serum and urinary angiotensinogen levels as prognostic indicators in acute kidney injury: a prospective study

doi: 10.1590/1806-9282.20230716

Figure Lengend Snippet: Correlation between urinary angiotensinogen/urine creatinine and serum angiotensinogen values.

Article Snippet: Before the analysis, the frozen samples were thawed at 2–8°C for 8 h. The sAGT and uAGT levels were measured using Human Angiotensinogen kits (Elabscience Biotechnology Co., Wuhan, PRC) and the enzyme-linked immunosorbent assay (ELISA) method in the biochemistry research laboratory of our hospital.

Techniques:

Journal: Revista da Associação Médica Brasileira

Article Title: Serum and urinary angiotensinogen levels as prognostic indicators in acute kidney injury: a prospective study

doi: 10.1590/1806-9282.20230716

Figure Lengend Snippet: Serum angiotensinogen and urinary angiotensinogen/urine creatinine ratio by stage of Kidney Disease: Improving Global Outcomes.

Article Snippet: Before the analysis, the frozen samples were thawed at 2–8°C for 8 h. The sAGT and uAGT levels were measured using Human Angiotensinogen kits (Elabscience Biotechnology Co., Wuhan, PRC) and the enzyme-linked immunosorbent assay (ELISA) method in the biochemistry research laboratory of our hospital.

Techniques:

Journal: Revista da Associação Médica Brasileira

Article Title: Serum and urinary angiotensinogen levels as prognostic indicators in acute kidney injury: a prospective study

doi: 10.1590/1806-9282.20230716

Figure Lengend Snippet: Relationship between urinary angiotensinogen/urine creatinine and hemodialysis and mortality.

Article Snippet: Before the analysis, the frozen samples were thawed at 2–8°C for 8 h. The sAGT and uAGT levels were measured using Human Angiotensinogen kits (Elabscience Biotechnology Co., Wuhan, PRC) and the enzyme-linked immunosorbent assay (ELISA) method in the biochemistry research laboratory of our hospital.

Techniques:

Journal: Experimental physiology

Article Title: Upregulation of a local renin-angiotensin system in the rat carotid body during chronic intermittent hypoxia.

doi: 10.1113/expphysiol.2013.074591

Figure Lengend Snippet: Figure 1. Expression and localization of angiotensinogen in the carotid body A, RT-PCR analysis of the mRNA expression of angiotensinogen (AGT) in the rat carotid body (CB). Aa, gel image of the RT-PCR products in normoxic (Nx) control and hypoxic (IH) groups. The expected size of PCR products is 312 bp from AGT and 436 bp from β-actin. Ab, the relative expression of the AGT/β-actin mRNA level (expressed as a percentage of the control value). Columns show mean values + SEM (n = 6 for each group). ∗P ≤0.05 versus the corresponding Nx control. B, in situ hybridization of AGT mRNA using digoxigenin-labelled antisense and sense riboprobes on cryosections of the CB. Ba, intense hybridization signals for AGT mRNA were detected in clusters of glomus cells in the IH group using antisense riboprobes. Bb, no positive hybridization signal for AGT mRNA was detected with sense riboprobes in IH. Bc, mild hybridization signals for AGT mRNA were detected in clusters of glomus cells in the Nx control. Bd, no positive hybridization signal for AGT mRNA was detected with sense riboprobes in Nx. Scale bar represents 40 μm

Article Snippet: Sections were incubated with primary antibodies to the following proteins: AGT (mouse monoclonal IgG antibody, 1:250 dilution; catalogue no. 10R-A131a; Fitzgerald Industries International, Acton, MA, USA) and AT1 (rabbit polyclonal IgG antibody, 1:250 dilution; catalogue no. sc-1173; Santa Cruz Biotechnology, Santa Cruz, CA, USA), in 0.05 M Tris–HCl buffer containing 2% bovine serum albumin at 4◦C overnight.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Control, In Situ Hybridization, Hybridization