goat α human ace2 ab  (R&D Systems)

Journal: Frontiers in Gastroenterology

Article Title: Alterations in serum and intestinal ACE2 in Inflammatory Bowel Disease and the impact of inflammation

doi: 10.3389/fgstr.2025.1590646

Figure Lengend Snippet: Corresponding H&E and ACEZ stained sections of (A) Healthy mouse lleum (Healthy mouse colon and (C) DSS mouse colon (n-5), magnification 20X The ileum and colon of healthy mice (A, B) and colon of Ds exposed mice (C) were stained with haematoxylin and eosin (H&E) for assessment of histological activity. H&E sections are represented on the left of the figure with the corresponding sections stained with ACE2 on the right.

Article Snippet: Sections were incubated overnight with ACE2 Ab (1:125, Santa Cruz, SC- 390851).

Techniques: Staining, Activity Assay

Journal: Frontiers in Gastroenterology

Article Title: Alterations in serum and intestinal ACE2 in Inflammatory Bowel Disease and the impact of inflammation

doi: 10.3389/fgstr.2025.1590646

Figure Lengend Snippet: lmmunohistochemical pattern of ACE2 staining in the terminal ileum and colon of healthy controls without evidence of intestinalinflammation. Intestinal tissue biopsies (terminal ileum and colon) were stained with anti ACE2 Ab (1:100). The predominant form of staining (brush border)is demonstrated.

Article Snippet: Sections were incubated overnight with ACE2 Ab (1:125, Santa Cruz, SC- 390851).

Techniques: Staining

Journal: Frontiers in Gastroenterology

Article Title: Alterations in serum and intestinal ACE2 in Inflammatory Bowel Disease and the impact of inflammation

doi: 10.3389/fgstr.2025.1590646

Figure Lengend Snippet: Corresponding H&E and ACE2 stained sections of (A) Healthy control ileaImucosa; (B) Inflamed Crohn's Disease ileal mucosa; (C) healthy control colonic mucosa and (D) Inflamed Ulcerative Colitis colon mucosa.The ileal and colonic mucosa of healthy controls and IBD patients were stained with haematoxylin and eosin (H&E) for assessment of histological activity. H&E sections are represented on the left of the figure with the corresponding sections stained with ACE2 on the right.

Article Snippet: Sections were incubated overnight with ACE2 Ab (1:125, Santa Cruz, SC- 390851).

Techniques: Staining, Control, Activity Assay

Journal: Frontiers in Gastroenterology

Article Title: Alterations in serum and intestinal ACE2 in Inflammatory Bowel Disease and the impact of inflammation

doi: 10.3389/fgstr.2025.1590646

Figure Lengend Snippet: Serum soluble ACE2levels (ng/ml)in Crohns Disease, Ulcerative Col tis and Healthy control cohorts.Serum ACE2 levels were measured by ELISA.

Article Snippet: Sections were incubated overnight with ACE2 Ab (1:125, Santa Cruz, SC- 390851).

Techniques: Control, Enzyme-linked Immunosorbent Assay

Journal: Journal of Virology

Article Title: Sensing of SARS-CoV-2-infected cells by plasmacytoid dendritic cells is modulated via an interplay between CD54/ICAM-1 and CD11a/LFA-1 α L integrin

doi: 10.1128/jvi.01235-24

Figure Lengend Snippet: Sensing of SARS-CoV-2-infected cells by pDCs is more efficient compared with cell-free virions. ( A ) Quantification of IFN-I (IFN-α/β) in the supernatant of PBMCs that were co-cultured for 24 h with SARS-CoV-2 (LSPQ1)-infected HeLa-hACE2, exposed to cell-free SARS-CoV-2 at different MOIs, or treated with supernatant of infected cells (MOI 0.1) (sup) collected at 24 h post-infection. PBMCs used in the co-cultures and in “cell-free virus exposure” conditions were from the same donors with the same number of cells. In the latter conditions, the MOI was based on the number of PBMCs. UI, uninfected cells. Each dot represents independent experiments/donors. Mann-Whitney U test. ** P < 0.01. ( B ) (Upper panel) Frequency of SARS-CoV-2 infection in HeLa-hACE2 as determined by flow cytometry using αN Ab and a mouse IgG1 isotype control (ctrl). (Lower panel) A summary dot plot shows the results from six independent infections. ( C ) Quantification of IFN-I in the supernatant of SARS-CoV-2-infected HeLa-hACE2 (MOI 0.1, 24 h) that were co-cultured with pDC-depleted PBMCs or total PBMCs ( n = 3 donors). ( D ) The analysis was as described for panel C but with cell-free viruses instead of infected cells ( n = 2 donors). The MOI (0.1) used was based on the number of PBMCs, as done in . In all relevant panels, the co-cultures were done for 24 h. Shown is the mean ± SEM. The dotted line indicates the limit of detection (LOD) of the assay (60 U/mL). See also .

Article Snippet: For the detection of the hACE2 receptor, cells were stained with 4 μg/mL of goat α-human ACE2 Ab (R&D system) or with the isotype control (goat IgG, R&D system) for 30 min at 4°C, followed by incubation with a secondary Ab.

Techniques: Infection, Cell Culture, Virus, MANN-WHITNEY, Flow Cytometry, Control

Journal: Journal of Virology

Article Title: Sensing of SARS-CoV-2-infected cells by plasmacytoid dendritic cells is modulated via an interplay between CD54/ICAM-1 and CD11a/LFA-1 α L integrin

doi: 10.1128/jvi.01235-24

Figure Lengend Snippet: pDC sensing of SARS-CoV-2-infected cells requires physical contact. ( A ) (Left panel) Schematic representation of the experimental design (created with BioRender.com). HeLa-hACE2 cells were infected with SARS-CoV-2 at MOI of 0.1 for 24 h and then co-cultured with PBMCs in the presence or absence of a 0.4 µm transwell membrane insert. PBMCs were added to the upper chamber of the insert. IFN-I was measured from the lower chamber, at 18–24 h after the co-cultures. (Right panel) Quantification of IFN-I in the supernatant of PBMCs co-cultured with mock- or SARS-CoV-2-infected HeLa-hACE2 (Inf cells) in the presence (+TW) or absence (–TW) of a transwell insert. Inf cells, infected cells. Data shown are mean ± SEM from independent experiments with seven donors. Each dot depicts a donor. Mann-Whitney U test. ** P < 0.01. ( B ) (Upper panel) Design of an experiment aimed at determining whether cell-free virus can trigger residual IFN-I production in the absence of physical contact (created with BioRender.com). (Lower panel) IFN-I was measured in supernatants of PBMCs either co-cultured with infected HeLa-hACE2 (MOI 0.1) in the presence or absence of transwell inserts or exposed to cell-free virus (MOI 0.1, based on PBMC number) in the same conditions. The number of PBMCs used for the co-cultures or exposed to cell-free viruses was the same. Data shown are mean ± SEM from independent experiments with four donors. Mann-Whitney U test. * P < 0.05. In both panels: LOD, limit of detection of the IFN-I assay as described in legend. See also .

Article Snippet: For the detection of the hACE2 receptor, cells were stained with 4 μg/mL of goat α-human ACE2 Ab (R&D system) or with the isotype control (goat IgG, R&D system) for 30 min at 4°C, followed by incubation with a secondary Ab.

Techniques: Infection, Cell Culture, Membrane, MANN-WHITNEY, Virus

Journal: Journal of Virology

Article Title: Sensing of SARS-CoV-2-infected cells by plasmacytoid dendritic cells is modulated via an interplay between CD54/ICAM-1 and CD11a/LFA-1 α L integrin

doi: 10.1128/jvi.01235-24

Figure Lengend Snippet: CD54/CD11a adhesion complex is involved in the recognition and sensing of SARS-CoV-2 infected cells by pDCs in a potentially bidirectional manner ( A ) Design of a CD54 blocking experiment (created with BioRender.com). PBMCs, enriched pDCs, or SARS-CoV-2-infected cells were pretreated with various concentrations of αCD54 Ab for 30 min at room temperature before the 24 h co-cultures. Cells treated with an isotype Ab control served as a negative control. ( B ) IFN-I in the supernatant of a co-culture between infected HeLa-hACE2 and αCD54 Ab-pretreated PBMCs. The IFN-I level in the Ab-untreated condition was set at 100%, six donors. ( C ) IFN-I production in the supernatant of a 24 h co-culture whereby infected HeLa-hACE2 were pretreated with αCD54 Ab and then exposed to PBMCs. The IFN-I level in the untreated condition was set at 100%, six donors. ( D ) IFN-I release from the supernatant of SARS-CoV-2-infected Calu-3 co-cultured with anti-CD54 Ab-pretreated PBMCs. Data analysis was as described for panel B, four donors. ( E ) IFN-I from a co-culture supernatant of PBMCs with infected Calu-3 that were treated with αCD54 Ab. Data analysis was as described for Panel B, four donors. ( F ) Experimental design of a CD11a blocking experiment (created with BioRender.com). Either PBMCs were pretreated with increasing concentrations of αCD11a Ab or the αCD11a Ab was added to a co-culture of PBMCs and infected Calu-3. IFN-I was determined 18–24 h post-co-culture. ( G and H ) IFN-I production in the supernatant of infected-Calu-3 co-cultured with PBMCs. ( G ) PBMCs were pretreated with the indicated concentrations of αCD11a Ab or isotype Ab control, and IFN-I release from a co-culture with untreated PBMCs was set at 100%. ( H ) αCD11a Ab was added to the co-culture of PBMCs and infected Calu-3. IFN-I production from the Ab-untreated co-culture was set as 100%, six donors. In all relevant panels, data are shown as mean ± SEM. Statistical analysis: Mann-Whitney U test; * P < 0.05, ** P < 0.01, ns, not significant. See also .

Article Snippet: For the detection of the hACE2 receptor, cells were stained with 4 μg/mL of goat α-human ACE2 Ab (R&D system) or with the isotype control (goat IgG, R&D system) for 30 min at 4°C, followed by incubation with a secondary Ab.

Techniques: Infection, Blocking Assay, Control, Negative Control, Co-Culture Assay, Cell Culture, MANN-WHITNEY

Journal: Journal of Virology

Article Title: Sensing of SARS-CoV-2-infected cells by plasmacytoid dendritic cells is modulated via an interplay between CD54/ICAM-1 and CD11a/LFA-1 α L integrin

doi: 10.1128/jvi.01235-24

Figure Lengend Snippet: Formation of conjugates between pDCs and SARS-CoV-2-infected cells is dependent on receptor engagement with CD54. ( A ) Schematic representation of the experimental design (created with BioRender.com). Mock- and SARS-CoV-2-infected HeLa-hACE2 were pre-labeled with cell-membrane dye eFluo 670 before co-culturing with PBMCs. Cell conjugates were analyzed by flow cytometry and defined as CD3 - CD14 - CD19 - ILT7 + BDCA2 + eFluor670 + population. Analysis was done 18–24 h after the co-culture. ( B ) (Upper panel) Dot plots showing the gating strategy to identify cell conjugates between pDCs (gated from PBMCs) and mock- or SARS-CoV-2-infected HeLa-hACE2. Numbers shown in the dot plots are cell percentages. (Lower panel) Frequency of cell conjugates from different experiments done with five donors. ( C ) (Upper panel) Representative dot plots showing the frequency of conjugates formed between enriched pDCs (CM-Dil + ) and mock- or SARS-CoV-2-infected HeLa-hACE2 (eFluor 670 + ). (Lower panel) Overall data from different experiments done with 7 PBMC donors. ( D ) (Upper panel) Representative dot plots showing the effect of blocking CD54 receptor engagement on conjugate formation between pDCs and infected HeLa-hACE2. Numbers indicated represent the percentage of conjugates. (Lower panel) Frequencies of conjugates between pDCs and eFluor670-labelled mock- and SARS-CoV-2-infected HeLa-hACE2 in the presence of αCD54 Ab or isotype control (5 µg/mL). Analysis was done with four donors. In all relevant panels, each line depicts a donor. Statistical analysis Mann-Whitney U test. * P < 0.05. See also .

Article Snippet: For the detection of the hACE2 receptor, cells were stained with 4 μg/mL of goat α-human ACE2 Ab (R&D system) or with the isotype control (goat IgG, R&D system) for 30 min at 4°C, followed by incubation with a secondary Ab.

Techniques: Infection, Labeling, Membrane, Flow Cytometry, Co-Culture Assay, Blocking Assay, Control, MANN-WHITNEY

Journal: ImmunoHorizons

Article Title: Human ACE2 Gene Replacement Mice Support SARS-CoV-2 Viral Replication and Nonlethal Disease Progression.

doi: 10.4049/immunohorizons.2400030

Figure Lengend Snippet: FIGURE 1. The full human ACE2 gene

Article Snippet: For analysis of ACE2 expression, tissues were stained with anti-ACE2 Ab (MAB933, R&D Systems), and binding detected using an ARK peroxidase kit (K3954, Dako), using 3,3-diaminobenzidine (DAB) as substrate.

Techniques:

Journal: ImmunoHorizons

Article Title: Human ACE2 Gene Replacement Mice Support SARS-CoV-2 Viral Replication and Nonlethal Disease Progression.

doi: 10.4049/immunohorizons.2400030

Figure Lengend Snippet: FIGURE 2. Human ACE2 and ACE2-DT expression in ACE2-GR mice.

Article Snippet: For analysis of ACE2 expression, tissues were stained with anti-ACE2 Ab (MAB933, R&D Systems), and binding detected using an ARK peroxidase kit (K3954, Dako), using 3,3-diaminobenzidine (DAB) as substrate.

Techniques: Expressing

Journal: ImmunoHorizons

Article Title: Human ACE2 Gene Replacement Mice Support SARS-CoV-2 Viral Replication and Nonlethal Disease Progression.

doi: 10.4049/immunohorizons.2400030

Figure Lengend Snippet: FIGURE 3. In situ human ACE2 protein expression in ACE2-GR mice.

Article Snippet: For analysis of ACE2 expression, tissues were stained with anti-ACE2 Ab (MAB933, R&D Systems), and binding detected using an ARK peroxidase kit (K3954, Dako), using 3,3-diaminobenzidine (DAB) as substrate.

Techniques: In Situ, Expressing

Journal: Viruses

Article Title: SARS-CoV-2 Modulation of HIV Latency Reversal in a Myeloid Cell Line: Direct and Bystander Effects

doi: 10.3390/v16081310

Figure Lengend Snippet: ( A ) Relative ACE2 expression levels in different U1 and U937 cells ( left ) and representative histograms of median fluorescence intensity (MFI) for each condition ( right ). Among U1, different conditions were evaluated, such as US: unstimulated U1; cfv: U1 exposed to SARS-CoV-2 cell-free; U937: unstimulated U937. U1 exposed to conditioned media (CM) obtained from non-infected MDM (Ni), MDM exposed to SARS-CoV-2 (Wh), MDM polarized (M1, or M2). PMA (Phorbol 12-myristate 13-acetate): positive control for reversal on both HIV-latently infected cell models. Data show the mean ± SEM, and statistical significance was calculated by one-way ANOVA. ( B ) Kinetics of the SARS-CoV-2 (Wh variant, MOI: 0.1) replication in myeloid (U1, U937) cells measured using RT-qPCR targeted to N and ORF1a genes in culture supernatant, as described in M&M. T0: supernatant obtained after three cell washes. 72 h: 3 days post-infection. Data are expressed as mean ± SD obtained from 4 independent experiments. (* p < 0.05; *** p < 0.001).

Article Snippet: A rabbit anti-human ACE2 primary polyclonal Ab (ab272690, Abcam, Cambridge, UK) and goat anti-rabbit IgG secondary (PE) (Abcam, UK) were used for ACE2 quantification.

Techniques: Expressing, Fluorescence, Infection, Positive Control, Variant Assay, Quantitative RT-PCR