Journal: Nano Today

Article Title: SARS-CoV-2 multi-variant rapid detector based on graphene transistor functionalized with an engineered dimeric ACE2 receptor

doi: 10.1016/j.nantod.2022.101729

Figure Lengend Snippet: Probing the interactions of ACE2 and Antibodies with SARS-CoV-2 Spike. A) Schematic representation of ACE2-mediated host cell entry mechanism. B) Cryo-EM structure of dimeric ACE2 receptor (the two monomers are colored differently). The two subunits of a monomer are reported: Peptidase domain (PD) and Collectrin-like domain (CLD) that is composed of neck domain (ND) and transmembrane α-helix (TM α-helix). C) Cryo-EM structure of soluble trimeric Spike protein (the three monomers have different colors). A zoomed-in view of the RBD is shown. D) Peptidase Domain of the ACE2 receptor bound to the RBD of the SARS-CoV-2 Spike protein. E) Antibody Anti-Spike CR3022 (Ab-CR3022), bound to the SARS-CoV-2 Spike protein RBD. F) Force profiles from the Steered Molecular Dynamics (SMD) simulations of RBD unbinding from the ACE2 receptor (orange) and Ab-CR3022 (green). G) Pull-down assay of Spike and ACE2 (upper), and Spike and Ab-CR3022 (lower). Control is represented by the same experiment excluding the Spike protein (bait) from the system. The binding of Spike with ACE2 or Ab-CR3022 were monitored by western blot analysis.

Article Snippet: Finally, the PBASE-functionalized devices were exposed to 20 µL of 250 μg/mL of anti-Spike (40150-R007; Sino Biological, Inc., China), ACE2 (10108-H08B-100; Sino Biological, Inc., China) or ACE2-Fc (Z03484–1; GenScript Biotech) separately and left overnight in a humidified environment at 4 °C.

Techniques: Cryo-EM Sample Prep, Pull Down Assay, Binding Assay, Western Blot

Journal: Nano Today

Article Title: SARS-CoV-2 multi-variant rapid detector based on graphene transistor functionalized with an engineered dimeric ACE2 receptor

doi: 10.1016/j.nantod.2022.101729

Figure Lengend Snippet: gFET setup and Spike recognition. A) gFET (size 10 mm × 10 mm) is composed of two source electrodes each one connected with six graphene channels and the respective drains. A single gate electrode is used for both sides of gFET. A schematic representation of the PBASE-modified gFET is reported in the inset panel. B) Raman Spectra of gFET (black) and gFET-PBASE (red) (diode laser wavelength 523 nm and laser power 50 mW). C) Schematic representation of gFET modified with ACE2. D-E) Detail of I ds -V g curves obtained for (D) ACE2_gFET (black) and ACE2_gFET + Spike (2 µg/mL) (red). E) ACE2_gFET (black) and ACE2_gFET + mPRO (2 µg/mL) (red). F) Comparative bar charts of ACE2_gFET before (black bars) and after (red bars) the addition of Spike (2 and 0.2 µg/mL) or mPRO (2 µg/mL). G) Schematic representation of gFET modified with Ab (Ab-CR3022). H-I) Detail of I ds -V g curves for (H) Ab_gFET (black) and Ab_gFET + Spike (2 µg/mL) (red); (I) Ab_gFET (black) and Ab_gFET + mPRO (2 µg/mL) (red); (J) Comparative bar charts of Ab_gFET before (black bars) and after (red bars) the addition of Spike (2, 0.2 and 0.02 µg/mL) or mPRO (2 µg/mL). Details of V g from 0 to 1.20 V are shown. In (F) and (J): * ** p < 0.001, * p < 0.05, error bars represent standard deviation (s.d.).

Article Snippet: Finally, the PBASE-functionalized devices were exposed to 20 µL of 250 μg/mL of anti-Spike (40150-R007; Sino Biological, Inc., China), ACE2 (10108-H08B-100; Sino Biological, Inc., China) or ACE2-Fc (Z03484–1; GenScript Biotech) separately and left overnight in a humidified environment at 4 °C.

Techniques: Modification, Standard Deviation

Journal: Nano Today

Article Title: SARS-CoV-2 multi-variant rapid detector based on graphene transistor functionalized with an engineered dimeric ACE2 receptor

doi: 10.1016/j.nantod.2022.101729

Figure Lengend Snippet: MD simulations of full length, soluble and Fc tagged ACE2 dimers. A) Representative structure of the full-length ACE2 dimer embedded in a membrane. B) Same as (A) but the membrane is omitted to show the TM helices. C) Soluble truncated ACE2 conformation. D) Starting configuration of the ACE2-Fc chimera. For B-D two orthogonal views are shown. E) Representative snapshots of ACE2-Fc structures sampled during the MD trajectory, side view. The PD centers of mass distance is shown by a dashed red line. F) Top view of soluble ACE2, the PD centers of mass distance is shown by a dashed black line. G) Number of contacts between the two PD-CLD regions of monomers for membrane embedded full length ACE2 (in black), ACE2-Fc (red) and soluble ACE2 (grey). H) Time evolution of the intermonomer distance measured between the PD domains, color code as in (G). Comparative Dynamics as reported by the cross-correlation matrices of concerted motions of the residues of our three dimeric systems during the MD simulations. In (I) correlations in the full length ACE2 embedded in the membrane (upper triangle) and soluble truncated ACE2 (lower triangle). L) Same comparison as (I) between full length ACE2 embedded in the membrane (upper triangle) and ACE2-Fc (lower triangle).

Article Snippet: Finally, the PBASE-functionalized devices were exposed to 20 µL of 250 μg/mL of anti-Spike (40150-R007; Sino Biological, Inc., China), ACE2 (10108-H08B-100; Sino Biological, Inc., China) or ACE2-Fc (Z03484–1; GenScript Biotech) separately and left overnight in a humidified environment at 4 °C.

Techniques:

Journal: Nano Today

Article Title: SARS-CoV-2 multi-variant rapid detector based on graphene transistor functionalized with an engineered dimeric ACE2 receptor

doi: 10.1016/j.nantod.2022.101729

Figure Lengend Snippet: ACE2-Fc as bioreceptor and LOD calculation. A) SDS-PAGE under reducing (w β lanes) and non-reducing (w/o β lanes) conditions of soluble ACE2 and ACE2-Fc; B) Pull-down assay of Spike and ACE2-Fc. The binding of Spike with ACE2-Fc was monitored by western blot. C) Schematic representation of gFET modified with ACE2-Fc. D) Comparative bar chart showing the ACE2-Fc_gFET response to different concentrations of SARS-CoV-2 Spike. E) Comparative bar chart showing the ACE2-Fc_gFET response to MERS-CoV Spike protein (2 µg/mL) and SARS-CoV-2 mPRO (2 µg/mL). In (D) and (E): * ** p < 0.001, * * p < 0.01 and * p < 0.05, error bars re p resent s.d.

Article Snippet: Finally, the PBASE-functionalized devices were exposed to 20 µL of 250 μg/mL of anti-Spike (40150-R007; Sino Biological, Inc., China), ACE2 (10108-H08B-100; Sino Biological, Inc., China) or ACE2-Fc (Z03484–1; GenScript Biotech) separately and left overnight in a humidified environment at 4 °C.

Techniques: SDS Page, Pull Down Assay, Binding Assay, Western Blot, Modification

Journal: Nano Today

Article Title: SARS-CoV-2 multi-variant rapid detector based on graphene transistor functionalized with an engineered dimeric ACE2 receptor

doi: 10.1016/j.nantod.2022.101729

Figure Lengend Snippet: POC device detects SARS-CoV-2 variants in clinical samples up to low viral concentrations. A) Detailed amino acid mutations of SARS-CoV-2 Spike proteins in B.1.610, Alpha, Delta, Omicron (BA.1, BA.2, BA.4, BA.5 and BQ.1) variants, compared to the wild-type Whuan-Hu1 of SARS-CoV-2. Positions of mutations are shown both on a schematic domain representation of the protein and on the 3D model (PDB ID: 7DWZ). B) Photograph of the gFET Cartridge Unit and the Signal acquisition modules connected to form the entire POC. A reference dimension bar is reported. Also, a schematic representation of gFET modified with ACE2-Fc tested with different SARS-CoV-2 samples is shown. C) Bar graph reporting ACE2-Fc_gFET signal before (black) and after the addition of nasopharyngeal swab samples from patients (red). D) Comparative bar chart showing the ACE2-Fc_gFET response to different dilutions of Omicron BA.5 swab. Ct and cps/mL of each patient or dilution are shown in or , respectively. In (C) and (D): *** p < 0.001, ** p < 0.01 and * p < 0.05, error bars re p resent s.d.

Article Snippet: Finally, the PBASE-functionalized devices were exposed to 20 µL of 250 μg/mL of anti-Spike (40150-R007; Sino Biological, Inc., China), ACE2 (10108-H08B-100; Sino Biological, Inc., China) or ACE2-Fc (Z03484–1; GenScript Biotech) separately and left overnight in a humidified environment at 4 °C.

Techniques: Modification

Journal: Diagnostics

Article Title: Postmortem Cardiopulmonary Pathology in Patients with COVID-19 Infection: Single-Center Report of 12 Autopsies from Lausanne, Switzerland

doi: 10.3390/diagnostics11081357

Figure Lengend Snippet: Antibodies used for immunofluorescence staining.

Article Snippet: ACE2 , CL4035 , Membranous , Mouse , Atlas Antibodies/ AMAB91262 , 1:1000.

Techniques: Immunofluorescence, Staining

Journal: Diagnostics

Article Title: Postmortem Cardiopulmonary Pathology in Patients with COVID-19 Infection: Single-Center Report of 12 Autopsies from Lausanne, Switzerland

doi: 10.3390/diagnostics11081357

Figure Lengend Snippet: Composition of antibodies used in the multiplexed immunofluorescence panels.

Article Snippet: ACE2 , CL4035 , Membranous , Mouse , Atlas Antibodies/ AMAB91262 , 1:1000.

Techniques: Immunofluorescence

Journal: Diagnostics

Article Title: Postmortem Cardiopulmonary Pathology in Patients with COVID-19 Infection: Single-Center Report of 12 Autopsies from Lausanne, Switzerland

doi: 10.3390/diagnostics11081357

Figure Lengend Snippet: ( A , B ) Multiplex imaging analysis depicting expression of ACE2, the major receptor for SARS-CoV-2, in different cellular compartments of lung tissues in a representative patient with COVID-19 (case #6). ( A ) Red arrows point to ACE2-expressing pneumocytes, defined by double-expression of ACE2 (yellow) and PANCK (white). ( B ) Cyan arrows highlight ACE2-expressing macrophages, defined by double-expression of ACE2 (yellow) and CD68 (red). All images were acquired on the Vectra Polaris microscope using a 20x objective magnification, and zoomed in areas are depicted for visualization of macrophages. ( C ) The was no difference in the relative frequencies (% of total imaged cells) of CD3+ T cells in lung control tissues (non-COVID-19 DAD, n = 5) and COVID-19 lung tissues ( n = 5), expressed as mean values with SD at the scatter dot plots (Mann–Whitney U test, p = 0.87). ( D ) There was no statistically significant difference of the CD4+ to CD8+ T lymphocyte ratio between the two cohorts (Mann–Whitney U test, p = 0.55).

Article Snippet: ACE2 , CL4035 , Membranous , Mouse , Atlas Antibodies/ AMAB91262 , 1:1000.

Techniques: Multiplex Assay, Imaging, Expressing, Microscopy, MANN-WHITNEY

Journal: Vaccines

Article Title: SARS-CoV-2 Spike Protein-Induced Interleukin 6 Signaling Is Blocked by a Plant-Produced Anti-Interleukin 6 Receptor Monoclonal Antibody

doi: 10.3390/vaccines9111365

Figure Lengend Snippet: SARS-CoV-2 spike-protein stimulation of human PBMCs and inhibition of spike-protein-induced IL-6 signaling. ( A ) PBMCs were incubated for 72 h with LPS (200 ng/mL, stimulation positive control) or recombinant SARS-CoV-2 S protein (1 µg/mL). The supernatant was collected, and IL-6 concentration was determined by ELISA. ( B ) Luciferase reporter cells were pre-incubated with various concentrations of pIL6RmAb or an IgG isotype antibody negative control (6.25 µg/mL). The supernatant from S protein-stimulated PBMCs was then added to the cells. After 24 h of incubation, Luciferase-detecting reagent was added and the bioluminescent signal was measured. The percentage of luciferase signal relative to no antibody treatment is shown. Student’s t-test and one-way ANOVA were used for statistical analysis. At least three independent experiments were performed, and error bars represent standard error of the mean. *** indicates p values < 0.001 of IL-6 secretion by the S protein treated PBMCs compared to that of no treatment negative control. * and ns (not significant) indicate p values < 0.05 and > 0.05, respectively, of pIL6RmAb-treated samples compared to that of no treatment negative control.

Article Snippet: Recombinant SARS-CoV-2 S1 protein (32-190005) was purchased from Abeomics (San Diego, CA, USA).

Techniques: Inhibition, Incubation, Positive Control, Recombinant, Concentration Assay, Enzyme-linked Immunosorbent Assay, Luciferase, Negative Control

Journal: BMC Cardiovascular Disorders

Article Title: Telmisartan protects chronic intermittent hypoxic mice via modulating cardiac renin-angiotensin system activity

doi: 10.1186/s12872-018-0875-4

Figure Lengend Snippet: TERT decreased the cardiac expression of ACE but increased the expression of ACE 2 and Ang II in CIH

Article Snippet: Rabbit-anti-mouse antibodies against ACE, ACE 2 and Ang II were obtained from the Wuhan Boster Company.

Techniques: Expressing, TUNEL Assay

Journal: BMC Cardiovascular Disorders

Article Title: Telmisartan protects chronic intermittent hypoxic mice via modulating cardiac renin-angiotensin system activity

doi: 10.1186/s12872-018-0875-4

Figure Lengend Snippet: ACE 2 staining in myocardial cells of mice from four experimental groups. a & b : CIH group; c & d :ARB group; e & f :blank control group; g & h : air control group. Immunohistochemistry was performed on the cardiac apex sections as described in Materials and methods. The expression level of cardiac ACE 2 was highest in ARB group followed by CIH group compared with the two control groups. Magnification: a , c , e and g : 100×; b , d , f and h : 400×. scale bar,100 μm

Article Snippet: Rabbit-anti-mouse antibodies against ACE, ACE 2 and Ang II were obtained from the Wuhan Boster Company.

Techniques: Staining, Immunohistochemistry, Expressing

Journal: Gastroenterology

Article Title: Human Intestinal Defensin 5 Inhibits SARS-CoV-2 Invasion by Cloaking ACE2

doi: 10.1053/j.gastro.2020.05.015

Figure Lengend Snippet: Bindings of peptides to ACE2. ( A ) Immunofluorescence displaying the locations of ACE2 ( red ) and FITC-HD5 ( green ) in human intestinal villus and enterocytes. The embedding graphs are the results of control groups. ( B ) Kinetics for HD5 RED binding to ACE2 loaded on AR2G biosensors activated by EDC and s-NHS. Fits of the data to a 1:1 binding model are shown with red dashes . ( C ) Kinetics for HD6 binding to ACE2 loaded on AR2G biosensors. ( D ) The deep free-energy well of HD5 docking onto the LBD of ACE2. The x-axis is the H-bond number. The y-axis is the root mean square deviation of backbone atoms of HD5. The z-axis is the free-energy landscape of HD5. ( E ) Energies of HD5 binding to ACE2 and SARS-CoV-2 S1-receptor-binding domain excluding the entropy effect.

Article Snippet: The bindings of HD5 and SARS-CoV-2 S1 to ACE2 (10108-H02H; Sino Biological) were measured using Forte Bio’s “Octet Red 96” BLI (Pall Life Sciences, New York, NY).

Techniques: Immunofluorescence, Control, Binding Assay

Journal: Gastroenterology

Article Title: Human Intestinal Defensin 5 Inhibits SARS-CoV-2 Invasion by Cloaking ACE2

doi: 10.1053/j.gastro.2020.05.015

Figure Lengend Snippet: HD5 binds to ACE2 and inhibits SARS-CoV-2 S1 binding and S pseudovirions entry to enterocytes. ( A ) Binding kinetics for HD5 and ACE2 loaded on streptavidin (SA) biosensors. Fits of the data to a 1:1 binding model are shown with red dashes. Times for association and dissociation are both 300 seconds. ( B ) Binding kinetics for SARS-CoV-2 S1 and ACE2 immobilized on SA biosensors. SARS-CoV-2 S1 is prepared in PBS with concentrations of 200, 100, 50, and 25 nM. ( C ) Biolayer interferometry-based ACE2 blocking experiment. The binding signals of 100 nM SARS-CoV-2 S1 to ACE2 coated with 600 nM HD5 are recorded for 120 seconds. ( D ) Stereoview of the cloak of HD5 on LBD. HD5 colored cyan is composed of 32 residues constrained by three disulfide bonds, displaying as a three-stranded antiparallel β-sheet conformation in steric. Residues in LBD (pink) cloaked by HD5 are colored black. ( E ) Immunofluorescence microscopy revealing the protection of HD5 on Caco-2 exposed to SARS-CoV-2 S1. SARS-CoV-2 S1 adhering to the cell surface is probed by a goat anti-rabbit Alexa Fluor 488 antibody (Green). Nuclei are stained using DAPI (blue). The embedding graph in sham group shows cells treated with HD5. The region of interest in SARS-CoV-2 S1-treated group is magnified in the embedding graph. ( F ) Luciferase assay. The experiment was conducted in triplicate and repeated three times in different days. Results are shown as mean ± standard deviation. Welch test indicated the differences among the groups excluding the sham group (F = 52.15, P = 1.51 × 10 –9 ). LSD test showed that, compared with the control group without HD5 treatment (n = 9; 148.2 (27.5%)), Caco-2 cells preincubated with HD5 for 1 h at concentrations of 10 μg/mL (n = 9; 68.88 (28.95 %); ∗∗∗∗, P = 3.23 × 10 –10 ), 50 μg/mL (n = 9; 2.25 (47.07 %); ∗∗∗∗, P = 2.89 × 10 –18 ), and 100 μg/mL (n = 9; 1.59 (37.69 %); ∗∗∗∗, P = 2.48 × 10 –18 ) were less sensitive to SARS-CoV-2 S pseudovirons invasion. The embedding graph shows the protein bands of SARS-CoV-2 S1 binding to Caco-2 treated with HD5. β-actin is the reference.

Article Snippet: The bindings of HD5 and SARS-CoV-2 S1 to ACE2 (10108-H02H; Sino Biological) were measured using Forte Bio’s “Octet Red 96” BLI (Pall Life Sciences, New York, NY).

Techniques: Binding Assay, Blocking Assay, Immunofluorescence, Microscopy, Staining, Luciferase, Standard Deviation, Control

Journal: Gastroenterology

Article Title: Human Intestinal Defensin 5 Inhibits SARS-CoV-2 Invasion by Cloaking ACE2

doi: 10.1053/j.gastro.2020.05.015

Figure Lengend Snippet: Western blot and immunofluorescence microscopy revealing the protection of HD5 on cells against SARS-CoV-2 invasion. ( A ) Western blot. Shown are the protein bands of SARS-CoV-2 S1 binding to Caco-2. HD5 preincubation had less of an effect on SARS-CoV-2 S1 binding to ACE2. ( B ) Immunofluorescence revealing the inhibition of HD5 on SARS-CoV-2 S pseudovirions entry to Caco-2 cells. The embedded graph in the sham group shows cells treated with HD5. The regions of interest in pseudovirions- and HD5-treated groups are magnified. ( C ) Schematic illustration of the HD5-mediated host innate defense against SARS-CoV-2. Paneth cell–secreted HD5 binds to ACE2 abundant on the intestinal epithelium, lowering viral entry by cloaking the LBD. ( D ) Inhibition of HD5 on SARS-CoV-2 S pseudovirions entry to human renal proximal tubular epithelial HK-2 cells. The embedded graph in the sham group shows cells treated with HD5.

Article Snippet: The bindings of HD5 and SARS-CoV-2 S1 to ACE2 (10108-H02H; Sino Biological) were measured using Forte Bio’s “Octet Red 96” BLI (Pall Life Sciences, New York, NY).

Techniques: Western Blot, Immunofluorescence, Microscopy, Binding Assay, Inhibition

Journal: Physiological Research

Article Title: Angiotensin I and II Stimulate Cell Invasion of SARS-CoV-2: Potential Mechanism via Inhibition of ACE2 Arm of RAS

doi: 10.33549/physiolres.935198

Figure Lengend Snippet: Characterization and use of HEK-ACE2 stable cell line. (a, b) Cytochemical and biochemical features of the model. The fluorescent photomicrographs of cells demonstrate the expression of ACE2 receptor on the cell surface (green: anti-ACE2 antibody, blue: nuclei stained with DAPI, scale bar: 20 μm) and Western blot analysis confirming the expression of ACE2. (c) Aa relative luminescence (RLU) recorded after incubation of HEK-ACE2 cells with PVPs for 48 h (MyBioSource) in serial dilutions (n=6). (d) The infectivity of PVPs produced in newly established cell line clones S/3 and S/7 in the cell-entry assay (n=5).

Article Snippet: To visualize the ACE2 expression in situ, the HEK-ACE2 cells were plated on cover glass pre-coated with rat-tail collagen, type I (Sigma-Aldrich), fixed with 4 % paraformaldehyde and treated with rabbit monoclonal antibody against human ACE2 at 10 μg/ml (Rockland Immunochemicals), followed by incubation with goat anti-rabbit IgG Alexa Fluor 488 secondary antibody (Invitrogen).

Techniques: Stable Transfection, Expressing, Staining, Western Blot, Incubation, Infection, Produced, Clone Assay

Journal: Physiological Research

Article Title: Angiotensin I and II Stimulate Cell Invasion of SARS-CoV-2: Potential Mechanism via Inhibition of ACE2 Arm of RAS

doi: 10.33549/physiolres.935198

Figure Lengend Snippet: The effect of Ang I and Ang II on PVP cell entry. The PVP infectivity (red line) was assessed using two cell lines (HEK-ACE2: a, b and Vero E6: c, d) at the indicated concentrations of peptides. The cell viability (blue line) was determined at the same concentrations of peptides in parallel cultures. Ang I and Ang II peptides stimulated the PVP entrance in HEK-ACE2 cells (red line, panel a, and b) at the concentration of 100 μM or higher (not shown), while the cell viability was not significantly decreased at the indicated concentrations (blue line). The increased infectivity of PVPs after the pre-treatment with Ang I (c) and Ang II (d) peptides in Vero E6 cells was observed; however, these cells showed decreased viability at the corresponding concentrations (blue line) (n=6, *** p<0.001; ** p<0.01; * p<0.05).

Article Snippet: To visualize the ACE2 expression in situ, the HEK-ACE2 cells were plated on cover glass pre-coated with rat-tail collagen, type I (Sigma-Aldrich), fixed with 4 % paraformaldehyde and treated with rabbit monoclonal antibody against human ACE2 at 10 μg/ml (Rockland Immunochemicals), followed by incubation with goat anti-rabbit IgG Alexa Fluor 488 secondary antibody (Invitrogen).

Techniques: Infection, Concentration Assay

Journal: Physiological Research

Article Title: Angiotensin I and II Stimulate Cell Invasion of SARS-CoV-2: Potential Mechanism via Inhibition of ACE2 Arm of RAS

doi: 10.33549/physiolres.935198

Figure Lengend Snippet: In vitro metabolism of Ang I and Ang II in HEK-ACE2 and Vero E6 cells in absence and presence of PVP. (a) The expected products after the Ang I and Ang II cleavage by RAS enzymes. (b–e) Quantification of angiotensin peptides in cell culture (n=3, ** p<0.01; * p<0.05) as determined by liquid chromatography and mass spectrometry (Details are in Material and Methods).

Article Snippet: To visualize the ACE2 expression in situ, the HEK-ACE2 cells were plated on cover glass pre-coated with rat-tail collagen, type I (Sigma-Aldrich), fixed with 4 % paraformaldehyde and treated with rabbit monoclonal antibody against human ACE2 at 10 μg/ml (Rockland Immunochemicals), followed by incubation with goat anti-rabbit IgG Alexa Fluor 488 secondary antibody (Invitrogen).

Techniques: In Vitro, Cell Culture, Liquid Chromatography, Mass Spectrometry

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