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: Frontiers in oncology

Article Title: The Angiotensin-Converting Enzyme Inhibitory State Promotes the Transformation of Non-Small Cell Lung Cancer Blood Supply Pattern Toward Vasculogenic Mimicry Formation.

doi: 10.3389/fonc.2021.663671

Figure Lengend Snippet: FIGURE 1 | High ACE2 expression was linked to increased VM and better prognosis in NSCLC. (A) Typical image of ACE2, VE-cadherin, EphA2 protein expression and CD34/PAS double staining in TMA tissues. Case B1 had massive CD34−/PAS+ VM (yellow arrows) lined by ACE2, VE-cadherin and EphA2 high expressing tumor cells. Case G1 had abundant CD34+/PAS−MVs (black arrows) with ACE2, VE-cadherin and EphA2 low expressing tumor cells. (B) Kaplan-Meier analysis of OS in NSCLC patients with ACE2 low or high expression. P = 0.044.

Article Snippet: The sections were dewaxed, rehydrated, treated with antigen retrieval, hatched with primary antibodies of ACE2 (1:200), VE-cadherin (1:200), EphA2 (1:100), and CD34 (1:200, ab81289, Abcam) overnight at 4°C, individually exposed to secondary antibody (SA1020, BOSTER, USA) 1 h at RT, and colored with DAB peroxidase substrate.

Techniques: Expressing, Double Staining

Journal: Frontiers in oncology

Article Title: The Angiotensin-Converting Enzyme Inhibitory State Promotes the Transformation of Non-Small Cell Lung Cancer Blood Supply Pattern Toward Vasculogenic Mimicry Formation.

doi: 10.3389/fonc.2021.663671

Figure Lengend Snippet: FIGURE 2 | ACE2-induced better outcome in NSCLC patients might be attributed to less vessels and more VM formation. (A, C, E, G) Typical tissue images of each group stained with ACE2 or CD34/PAS. Yellow arrows: CD34−/PAS+ VMs; black arrows: CD34+/PAS−MV. (B, D, F, H) Kaplan–Meier analysis of OS in each group.

Article Snippet: The sections were dewaxed, rehydrated, treated with antigen retrieval, hatched with primary antibodies of ACE2 (1:200), VE-cadherin (1:200), EphA2 (1:100), and CD34 (1:200, ab81289, Abcam) overnight at 4°C, individually exposed to secondary antibody (SA1020, BOSTER, USA) 1 h at RT, and colored with DAB peroxidase substrate.

Techniques: Staining

Journal: Frontiers in oncology

Article Title: The Angiotensin-Converting Enzyme Inhibitory State Promotes the Transformation of Non-Small Cell Lung Cancer Blood Supply Pattern Toward Vasculogenic Mimicry Formation.

doi: 10.3389/fonc.2021.663671

Figure Lengend Snippet: FIGURE 3 | Human ACE2 was stably overexpressed in A549-ACE2-OE cells. (A) Schematic representation of pLenti6.3-MCS/V5 DEST. (B) pLenti6.3-ACE2 expression vector was detected by PCR. (C) Fluorescence of EGFP in A549-ACE2-OE cells (left) and parental cells (right) was determined by fluorescence microscopy. (D) RT-PCR experiment of ACE2 mRNA level in A549-ACE2-OE cells and control cells, Mean ± SD, n = 3, *p < 0.05. (E) Western blot analysis of ACE2 expression level in A549-ACE2-OE cells and parental cells. (F) Quantification of ACE2 expression level in A549-ACE2-OE cells and parental cells. Mean ± SD, n = 3, ***p < 0.001.

Article Snippet: The sections were dewaxed, rehydrated, treated with antigen retrieval, hatched with primary antibodies of ACE2 (1:200), VE-cadherin (1:200), EphA2 (1:100), and CD34 (1:200, ab81289, Abcam) overnight at 4°C, individually exposed to secondary antibody (SA1020, BOSTER, USA) 1 h at RT, and colored with DAB peroxidase substrate.

Techniques: Stable Transfection, Expressing, Plasmid Preparation, Fluorescence, Microscopy, Reverse Transcription Polymerase Chain Reaction, Control, Western Blot

Journal: Frontiers in oncology

Article Title: The Angiotensin-Converting Enzyme Inhibitory State Promotes the Transformation of Non-Small Cell Lung Cancer Blood Supply Pattern Toward Vasculogenic Mimicry Formation.

doi: 10.3389/fonc.2021.663671

Figure Lengend Snippet: FIGURE 4 | Tube formation ability of A549 cells was improved with ACE inhibitory state. (A) Morphologies of a panel of A549-ACE2-OE cells, A549-NC cells, and A549-NC cells treated with ACEI (1, 5, and 10 nM/L) were shown as sheet-like and thread-like cell types in 2D culture, which were outlined partly with yellow lines. Representative images were shown above. Upper: white light; lower: fluorescence. (B) Quantification of sheet-like or thread-like cells and pebble-like cells in three groups, representatively, Mean ± SD, n = 3, **p < 0.01. (C) Images of both above cell lines grown in 3D Matrix gel. A 10 nM/L ACEI dilution was performed. Representative images were shown above. Yellow arrows point out the free cancer cells escaping from tube wall and isolated segments. Upper: white light; lower: fluorescence. (D, E) Images of 3D culture were applied to determine average number of nodes, branches, isolated segments, meshes and mean mesh area in those groups, per field, Mean ± SD, n = 3, **p < 0.01, *p < 0.05. ns, no significance.

Article Snippet: The sections were dewaxed, rehydrated, treated with antigen retrieval, hatched with primary antibodies of ACE2 (1:200), VE-cadherin (1:200), EphA2 (1:100), and CD34 (1:200, ab81289, Abcam) overnight at 4°C, individually exposed to secondary antibody (SA1020, BOSTER, USA) 1 h at RT, and colored with DAB peroxidase substrate.

Techniques: Isolation

Journal: Frontiers in oncology

Article Title: The Angiotensin-Converting Enzyme Inhibitory State Promotes the Transformation of Non-Small Cell Lung Cancer Blood Supply Pattern Toward Vasculogenic Mimicry Formation.

doi: 10.3389/fonc.2021.663671

Figure Lengend Snippet: FIGURE 5 | VM formation was increased, and vasculature was lessened due to inhibition of RAS in vivo. (A) Growth curve of allograft tumors of A549-ACE2-OE cells, A549-NC cells with or without ACEI treatment, Mean ± SD, n = 3, *p < 0.05. ns, no significance. (B) Weight of resected tumors, Mean ± SD, n = 3, **p < 0.01, *p < 0.05. (C) Continuous sections of allograft tumor tissues stained with PAS, CD34, VE-cadherin, or EphA2 immunohistochemical stain. Black arrow points out a typical MV (CD34+/PAS−); yellow arrows point out typical VM (CD34−/PAS+). (D) Quantification of MV and VM in different groups, Mean ± SD, n = 3, per field, ***p < 0.001, **p < 0.01. (E) Quantification of VE-cadherin and EphA2 mean optical density in three groups, Mean ± SD, n = 3, ***p < 0.001, **p < 0.01.

Article Snippet: The sections were dewaxed, rehydrated, treated with antigen retrieval, hatched with primary antibodies of ACE2 (1:200), VE-cadherin (1:200), EphA2 (1:100), and CD34 (1:200, ab81289, Abcam) overnight at 4°C, individually exposed to secondary antibody (SA1020, BOSTER, USA) 1 h at RT, and colored with DAB peroxidase substrate.

Techniques: Inhibition, In Vivo, Staining, Immunohistochemical staining

Journal: Frontiers in oncology

Article Title: The Angiotensin-Converting Enzyme Inhibitory State Promotes the Transformation of Non-Small Cell Lung Cancer Blood Supply Pattern Toward Vasculogenic Mimicry Formation.

doi: 10.3389/fonc.2021.663671

Figure Lengend Snippet: FIGURE 6 | VE-cadherin and EphA2 expression was upregulated in A549 cells and NSCLC tissues with impaired local RAS status. (A, B) RT-PCR experiment of VE-cadherin and EphA2 mRNA level in A549-ACE2-OE cells and control cells, Mean ± SD, n = 3, ***p < 0.001. (C–E) Western blot analysis and quantification of VE- cadherin and EphA2 expression level in A549-ACE2-OE cells and control cells, Mean ± SD, n = 3, ***p < 0.001. (F, G) Linear regressions of VM number and VE- cadherin (P < 0.0001) or EphA2 (P = 0.0108) score in TMA. (H) Typical tissue images of both groups stained with VE-cadherin, ACE2 or CD34/PAS. Case F13 with ACE2 low status was provided with rambling VM covered by tumor cells which only expressed VE-cadherin in nuclei; case D6 with ACE2 high status had ordered VM lined by tumor cells expressing VE-cadherin in both nuclei and cytomembranes. Red arrow: VE-cadherin membrane expression.

Article Snippet: The sections were dewaxed, rehydrated, treated with antigen retrieval, hatched with primary antibodies of ACE2 (1:200), VE-cadherin (1:200), EphA2 (1:100), and CD34 (1:200, ab81289, Abcam) overnight at 4°C, individually exposed to secondary antibody (SA1020, BOSTER, USA) 1 h at RT, and colored with DAB peroxidase substrate.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Western Blot, Staining, Membrane

Journal: Frontiers in oncology

Article Title: The Angiotensin-Converting Enzyme Inhibitory State Promotes the Transformation of Non-Small Cell Lung Cancer Blood Supply Pattern Toward Vasculogenic Mimicry Formation.

doi: 10.3389/fonc.2021.663671

Figure Lengend Snippet: FIGURE 7 | PI3K/AKT, p38MAPK, and HIF1-a were inactivated, and Nodal/Notch4 pathway was activated in A549-ACE2-OE cell model. (A–E) Western blot analysis and quantification of AKT, p-AKT, p38, and p-p38 expression level in A549-ACE2-OE cells and negative control, Mean ± SD, n = 3, ***p < 0.001, **p < 0.01. (F, G) Immunoflurescence assay and quantification of HIF1-a mean optical density in A549-ACE2-OE cells and negative control, Mean ± SD, n = 4, *p < 0.05. (H, I) Western blot analysis and quantification of Nodal and Notch4 expression level in A549-ACE2-OE cells and negative control, Mean ± SD, n = 3, ***p < 0.001, **p < 0.01.

Article Snippet: The sections were dewaxed, rehydrated, treated with antigen retrieval, hatched with primary antibodies of ACE2 (1:200), VE-cadherin (1:200), EphA2 (1:100), and CD34 (1:200, ab81289, Abcam) overnight at 4°C, individually exposed to secondary antibody (SA1020, BOSTER, USA) 1 h at RT, and colored with DAB peroxidase substrate.

Techniques: Western Blot, Expressing, Negative Control

Journal: Nature Communications

Article Title: A method for Boolean analysis of protein interactions at a molecular level

doi: 10.1038/s41467-022-32395-w

Figure Lengend Snippet: Antibodies used for MolBoolean experiments

Article Snippet: Mouse anti-ACE2 Clone CL4013 , 5 μg/mL , Atlas Antibodies, AMAb91259, 03083.

Techniques: Concentration Assay, Transduction

Journal: PLoS ONE

Article Title: Endoplasmic Reticulum Stress-Sensing Mechanism Is Activated in Entamoeba histolytica upon Treatment with Nitric Oxide

doi: 10.1371/journal.pone.0031777

Figure Lengend Snippet: Genes upregulated encoding metabolism enzymes.

Article Snippet: The primary antibody used were a rabbit anti-PFOR polyclonal antibody (1∶400 dilution, a kind gift from Dr Ester Orozco, CINVESTAV, Mexico), a mouse anti-actin monoclonal antibody (1∶20,000 dilution; #69100, MP Biomedicals).

Techniques:

Journal: PLoS ONE

Article Title: Endoplasmic Reticulum Stress-Sensing Mechanism Is Activated in Entamoeba histolytica upon Treatment with Nitric Oxide

doi: 10.1371/journal.pone.0031777

Figure Lengend Snippet: PFOR of 120 kDa was revealed upon SDS-PAGE and immunobloting of crude extracts from SNP-treated and control trophozoites (4, 2 and 1×10 4 cells). The loaded amount of proteins for each condition is evidenced by actin (43 kDa) immunodetection on the same western blot. Quantification of signal emission did not reveal differences between the tested conditions in 3 independent experiments.

Article Snippet: The primary antibody used were a rabbit anti-PFOR polyclonal antibody (1∶400 dilution, a kind gift from Dr Ester Orozco, CINVESTAV, Mexico), a mouse anti-actin monoclonal antibody (1∶20,000 dilution; #69100, MP Biomedicals).

Techniques: SDS Page, Western Blot, Control, Immunodetection

Journal: PLoS ONE

Article Title: Endoplasmic Reticulum Stress-Sensing Mechanism Is Activated in Entamoeba histolytica upon Treatment with Nitric Oxide

doi: 10.1371/journal.pone.0031777

Figure Lengend Snippet: PFOR activities upon no treatment.

Article Snippet: The primary antibody used were a rabbit anti-PFOR polyclonal antibody (1∶400 dilution, a kind gift from Dr Ester Orozco, CINVESTAV, Mexico), a mouse anti-actin monoclonal antibody (1∶20,000 dilution; #69100, MP Biomedicals).

Techniques: Activity Assay

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: 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