ace2 genes Search Results


86
Thermo Fisher gene exp ace2 hs01085333 m1
Gene Exp Ace2 Hs01085333 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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96
Sino Biological human ace2
Characterization of an <t>AAV6.2FF-hACE2</t> transduction model for SARS-CoV-2 infection of wild-type mice (A) Diagram of AAV genome expressing hACE2 from the CASI promoter. (B) Western blot of HEK293 cells transduced with AAV6.2FF-hACE and probed with an anti-hACE2 antibody. (C) BALB/c mice were administered 1 x 10 11 vg of AAV-Luc intranasally and imaged 10 days later using an IVIS imager. (D–F) (D) IFA images of lungs harvested from BALB/c mice infected intranasally with 1 x 10 11 vg of AAV-hACE2 or AAV-Luc and euthanized 10 days later. Lungs were stained with a rabbit anit-hACE2 antibody and imaged at 20 X (scale bar, 50mM). Viral RNA (E), and virus TCID50 titers (F) were determined in respiratory tissues on days 2 and 4 post-infection. n = 6 (3M, 3F). Statistical significance determined by Mann-Whitney test. ∗ = p < 0.05, ∗∗ = p < 0.01.
Human Ace2, supplied by Sino Biological, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ace2+genes/pmc08186956-212-3-5?v=Sino+Biological
Average 96 stars, based on 1 article reviews
human ace2 - by Bioz Stars, 2026-08
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87
Thermo Fisher gene exp ace2 rn01416293 m1
Figure 2. Renal and renal artery mRNA gene expression of angiotensin-converting enzyme (ACE), <t>ACE2,</t> neprilysin, angiotensin II receptor type 1a (AT1aR), angiotensin II receptor type 2 (AT2R), and Mas receptor (MasR) in Wistar Kyoto (WKY), WKY.SPGlaY, spontaneously hypertensive stroke-prone rats (SHRSP), and SP.WKYGlaY (n>5 per group). All relative gene expression data are expressed relative to the WKY group and are presented as mean±SEM. *P<0.05, **P<0.01, ***P<0.005 compared with WKY; #P<0.05, ##P<0.01 compared with SHRSP.
Gene Exp Ace2 Rn01416293 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 87/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ace2+genes/10__1161_slash_hypertensionaha__114__03756-33-9-3?v=Thermo+Fisher
Average 87 stars, based on 1 article reviews
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96
Sino Biological human ace2 cdna
Figure 2. Renal and renal artery mRNA gene expression of angiotensin-converting enzyme (ACE), <t>ACE2,</t> neprilysin, angiotensin II receptor type 1a (AT1aR), angiotensin II receptor type 2 (AT2R), and Mas receptor (MasR) in Wistar Kyoto (WKY), WKY.SPGlaY, spontaneously hypertensive stroke-prone rats (SHRSP), and SP.WKYGlaY (n>5 per group). All relative gene expression data are expressed relative to the WKY group and are presented as mean±SEM. *P<0.05, **P<0.01, ***P<0.005 compared with WKY; #P<0.05, ##P<0.01 compared with SHRSP.
Human Ace2 Cdna, supplied by Sino Biological, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ace2+genes/pm36457995-170-0-4?v=Sino+Biological
Average 96 stars, based on 1 article reviews
human ace2 cdna - by Bioz Stars, 2026-08
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91
Thermo Fisher gene exp ace2 hs01085334 m1
mRNA target selection from the 30 probes included in the prognostically relevant signature and TaqMan® gene expression assay description.
Gene Exp Ace2 Hs01085334 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ace2+genes/pmc03739775-31-28--1?v=Thermo+Fisher
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96
Sino Biological hek293t cells
Construction of <t>HEK293T</t> cells line that continuously expressing ACE2-GFP. HEK293Tcells were transfected with pCMV-ACE2-GFPSark tag plasmid and selected with hygromycin B to generate cell lines expressingACE2-GFP, cells were passaged 10 times to ensure stable expression. a Confocal imaging of HEK293T cell line with stably expressed ACE2-GFP. b Cell lysates were analyzed by Western blot with antibodies specific for ACE2 to confirm the expression of ACE2-GFP in HEK293T cells
Hek293t Cells, supplied by Sino Biological, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ace2+genes/pmc08387204-115-8-11?v=Sino+Biological
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96
Sino Biological human pcmv3 ace2 flag plasmid
Beas-2B cell migration was decreased by the treatment with SARS-CoV-2 (2019-nCoV) spike protein. (A, B) Beas-2B cells were transfected with pCMV control or <t>pCMV3-ACE2-Flag</t> plasmids DNA for 24 h. Cells were treated with 25 ng/mL SARS-CoV-2 (2019-nCoV) spike protein in the presence or absence of FBS for another 24 h. The cells were fixed and stained with antibody against Arl13b and DAPI. The ciliated cells were counted (n>500 cells) (A). Cell lysates were prepared and the protein expression of ACE2-Flag was detected by western blotting using antibody against Flag (B). (C-E) Beas-2B cells were plated on 35-mm 2 dishes and incubated for 24 h. A confluent monolayer of Beas-2B cells was then scratched with a sterile pipet tip. Then, cells were treated with 25 ng/mL spike protein and incubated for 24 h. Migration of cells into the space left by the scratch was photographed using a phase-contrast microscope at 200× magnification (C). Percentage of cell migration was quantified by subtracting the empty area remained at each time point from that at the initiation using NIH image analysis software (version 1.62; National Institutes of Health), and compared to that of the 0-h time point. Percentage of cell migration was presented as bar graph (D). Changes in single cell path were analyzed by tracking program, ImageJ plugin MTrackJ (Version 1.5.0). Movement track was presented in coordinates for each cell on a polar grid with the normalization of start points to the origin (E). Data in bar graphs represented as means ± SD. # p <0.05, significantly different from pCMV-transfected and spike protein-untreated group incubated with or without FBS & p <0.05, significantly different from pCMV3-ACE2-transfected and spike protein-untreated group incubated with or without FBS (A, D). ** p <0.01, significantly different from spike protein-untreated group (D).
Human Pcmv3 Ace2 Flag Plasmid, supplied by Sino Biological, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ace2+genes/pmc08902458-35-0-13?v=Sino+Biological
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96
Sino Biological plv c gfpspark hace2
BHK cells stably expressing GFP and <t>hACE2</t> were incubated with His-tagged MERS-CoV RBD, SARS-CoV-2 WT RBD, Alpha RBD, Beta RBD, Gamma RBD, Mink-Y453F RBD, Mink-N501T RBD, and Mink-F486L RBD, respectively. APC anti-His antibodies were used to detect the His-tagged protein binding to the cells. Representative results from three experiments are shown. The mean ± SD percentages of RBD-binding cells in the three experiments are shown in the right lower bar chart. Statistical significance was analyzed using one-way ANOVA with a Tukey’s multiple comparison test for multiple groups.
Plv C Gfpspark Hace2, supplied by Sino Biological, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ace2+genes/pmc08528823-186-9-10?v=Sino+Biological
Average 96 stars, based on 1 article reviews
plv c gfpspark hace2 - by Bioz Stars, 2026-08
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85
Thermo Fisher gene exp ace2 rn01416295 m1
BHK cells stably expressing GFP and <t>hACE2</t> were incubated with His-tagged MERS-CoV RBD, SARS-CoV-2 WT RBD, Alpha RBD, Beta RBD, Gamma RBD, Mink-Y453F RBD, Mink-N501T RBD, and Mink-F486L RBD, respectively. APC anti-His antibodies were used to detect the His-tagged protein binding to the cells. Representative results from three experiments are shown. The mean ± SD percentages of RBD-binding cells in the three experiments are shown in the right lower bar chart. Statistical significance was analyzed using one-way ANOVA with a Tukey’s multiple comparison test for multiple groups.
Gene Exp Ace2 Rn01416295 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ace2+genes/pmc03356861-86-9-10?v=Thermo+Fisher
Average 85 stars, based on 1 article reviews
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90
Thermo Fisher gene exp ace2 hs01085340 m1
The other does not include the transmembrane domain (pink) and lays nested between two motifs known to interact with the SARS-CoV receptor binding domain (blue). The measure of soluble <t>ACE2</t> represents normalized expression of the pink to yellow target, or the proportion of non-transmembrane ACE2 to transmembrane ACE2 .
Gene Exp Ace2 Hs01085340 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ace2+genes/med_rxiv__2020__11__23__20237206-60-19--1?v=Thermo+Fisher
Average 90 stars, based on 1 article reviews
gene exp ace2 hs01085340 m1 - by Bioz Stars, 2026-08
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96
Sino Biological c gfpspark tag
The other does not include the transmembrane domain (pink) and lays nested between two motifs known to interact with the SARS-CoV receptor binding domain (blue). The measure of soluble <t>ACE2</t> represents normalized expression of the pink to yellow target, or the proportion of non-transmembrane ACE2 to transmembrane ACE2 .
C Gfpspark Tag, supplied by Sino Biological, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ace2+genes/pmc09429913-104-6-16?v=Sino+Biological
Average 96 stars, based on 1 article reviews
c gfpspark tag - by Bioz Stars, 2026-08
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Image Search Results


Characterization of an AAV6.2FF-hACE2 transduction model for SARS-CoV-2 infection of wild-type mice (A) Diagram of AAV genome expressing hACE2 from the CASI promoter. (B) Western blot of HEK293 cells transduced with AAV6.2FF-hACE and probed with an anti-hACE2 antibody. (C) BALB/c mice were administered 1 x 10 11 vg of AAV-Luc intranasally and imaged 10 days later using an IVIS imager. (D–F) (D) IFA images of lungs harvested from BALB/c mice infected intranasally with 1 x 10 11 vg of AAV-hACE2 or AAV-Luc and euthanized 10 days later. Lungs were stained with a rabbit anit-hACE2 antibody and imaged at 20 X (scale bar, 50mM). Viral RNA (E), and virus TCID50 titers (F) were determined in respiratory tissues on days 2 and 4 post-infection. n = 6 (3M, 3F). Statistical significance determined by Mann-Whitney test. ∗ = p < 0.05, ∗∗ = p < 0.01.

Journal: iScience

Article Title: A novel mouse AAV6 hACE2 transduction model of wild-type SARS-CoV-2 infection studied using synDNA immunogens

doi: 10.1016/j.isci.2021.102699

Figure Lengend Snippet: Characterization of an AAV6.2FF-hACE2 transduction model for SARS-CoV-2 infection of wild-type mice (A) Diagram of AAV genome expressing hACE2 from the CASI promoter. (B) Western blot of HEK293 cells transduced with AAV6.2FF-hACE and probed with an anti-hACE2 antibody. (C) BALB/c mice were administered 1 x 10 11 vg of AAV-Luc intranasally and imaged 10 days later using an IVIS imager. (D–F) (D) IFA images of lungs harvested from BALB/c mice infected intranasally with 1 x 10 11 vg of AAV-hACE2 or AAV-Luc and euthanized 10 days later. Lungs were stained with a rabbit anit-hACE2 antibody and imaged at 20 X (scale bar, 50mM). Viral RNA (E), and virus TCID50 titers (F) were determined in respiratory tissues on days 2 and 4 post-infection. n = 6 (3M, 3F). Statistical significance determined by Mann-Whitney test. ∗ = p < 0.05, ∗∗ = p < 0.01.

Article Snippet: The cDNA for human ACE2 (SinoBiological; HG10108-M) was cloned into an AAV genome plasmid (pACASI-MSC-WPRE) containing the composite CASI promoter ( ) consisting of the human cytomegalovirus immediate-early gene enhancer region, the chicken beta actin promoter, and the human ubiquitin C promoter, as well as the woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) and simian virus 40 polyadenylation sequence downstream of hACE2 with flanking AAV2 inverted terminal repeats (ITRs).

Techniques: Transduction, Infection, Expressing, Western Blot, Staining, MANN-WHITNEY

SARS-CoV-2 spike DNA antigens protect from viral replication in vivo (A) Mice were immunized once or twice separated by four weeks with 10ug of pS via electroporation. Serum was collected at day 18 post-final immunization. At 35 days post-final immunization mice were infected intranasally with adeno-associated virus expressing human ACE2 (white). 17 days following AAV6-ACE2 transduction, animals were intranasally infected with 1 × 10 5 PFU of SARS-CoV-2 VIDO-01 P2. Four days post infection, animals were sacrificed to quantify viral replication. SARS-CoV-2 specific serum IgG endpoint titers (B) and pseudoviral neutralization titers (C) at day 18 post-final immunization. Replication competent virus (D), and viral RNA (E) in the lungs four-days post-infection. Pearson correlations between virus titer and serum IgG endpoints (F) and neutralization titers (G). Pearson correlations between viral RNA copies and serum IgG endpoints (H) and neutralization titers (I). Each point represents the average of duplicate samples from an individual animal, bars represent the mean, lines represent the median, and error bars represent the SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗∗p < 0.0001 by student's t-test (A and B), or Kruskall-Wallis ANOVA (D and E). Spearman correlations were used to determine relationships (F-I). Data are representative of one experiment with n = 5 males (squares) and 5 females (circles) per group.

Journal: iScience

Article Title: A novel mouse AAV6 hACE2 transduction model of wild-type SARS-CoV-2 infection studied using synDNA immunogens

doi: 10.1016/j.isci.2021.102699

Figure Lengend Snippet: SARS-CoV-2 spike DNA antigens protect from viral replication in vivo (A) Mice were immunized once or twice separated by four weeks with 10ug of pS via electroporation. Serum was collected at day 18 post-final immunization. At 35 days post-final immunization mice were infected intranasally with adeno-associated virus expressing human ACE2 (white). 17 days following AAV6-ACE2 transduction, animals were intranasally infected with 1 × 10 5 PFU of SARS-CoV-2 VIDO-01 P2. Four days post infection, animals were sacrificed to quantify viral replication. SARS-CoV-2 specific serum IgG endpoint titers (B) and pseudoviral neutralization titers (C) at day 18 post-final immunization. Replication competent virus (D), and viral RNA (E) in the lungs four-days post-infection. Pearson correlations between virus titer and serum IgG endpoints (F) and neutralization titers (G). Pearson correlations between viral RNA copies and serum IgG endpoints (H) and neutralization titers (I). Each point represents the average of duplicate samples from an individual animal, bars represent the mean, lines represent the median, and error bars represent the SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗∗p < 0.0001 by student's t-test (A and B), or Kruskall-Wallis ANOVA (D and E). Spearman correlations were used to determine relationships (F-I). Data are representative of one experiment with n = 5 males (squares) and 5 females (circles) per group.

Article Snippet: The cDNA for human ACE2 (SinoBiological; HG10108-M) was cloned into an AAV genome plasmid (pACASI-MSC-WPRE) containing the composite CASI promoter ( ) consisting of the human cytomegalovirus immediate-early gene enhancer region, the chicken beta actin promoter, and the human ubiquitin C promoter, as well as the woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) and simian virus 40 polyadenylation sequence downstream of hACE2 with flanking AAV2 inverted terminal repeats (ITRs).

Techniques: In Vivo, Electroporation, Infection, Expressing, Transduction, Neutralization

Journal: iScience

Article Title: A novel mouse AAV6 hACE2 transduction model of wild-type SARS-CoV-2 infection studied using synDNA immunogens

doi: 10.1016/j.isci.2021.102699

Figure Lengend Snippet:

Article Snippet: The cDNA for human ACE2 (SinoBiological; HG10108-M) was cloned into an AAV genome plasmid (pACASI-MSC-WPRE) containing the composite CASI promoter ( ) consisting of the human cytomegalovirus immediate-early gene enhancer region, the chicken beta actin promoter, and the human ubiquitin C promoter, as well as the woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) and simian virus 40 polyadenylation sequence downstream of hACE2 with flanking AAV2 inverted terminal repeats (ITRs).

Techniques: Recombinant, Plasmid Preparation, Enzyme-linked Immunospot, Software

Figure 2. Renal and renal artery mRNA gene expression of angiotensin-converting enzyme (ACE), ACE2, neprilysin, angiotensin II receptor type 1a (AT1aR), angiotensin II receptor type 2 (AT2R), and Mas receptor (MasR) in Wistar Kyoto (WKY), WKY.SPGlaY, spontaneously hypertensive stroke-prone rats (SHRSP), and SP.WKYGlaY (n>5 per group). All relative gene expression data are expressed relative to the WKY group and are presented as mean±SEM. *P<0.05, **P<0.01, ***P<0.005 compared with WKY; #P<0.05, ##P<0.01 compared with SHRSP.

Journal: Hypertension

Article Title: Origin of the Y Chromosome Influences Intrarenal Vascular Responsiveness to Angiotensin I and Angiotensin (1-7) in Stroke-Prone Spontaneously Hypertensive Rats

doi: 10.1161/hypertensionaha.114.03756

Figure Lengend Snippet: Figure 2. Renal and renal artery mRNA gene expression of angiotensin-converting enzyme (ACE), ACE2, neprilysin, angiotensin II receptor type 1a (AT1aR), angiotensin II receptor type 2 (AT2R), and Mas receptor (MasR) in Wistar Kyoto (WKY), WKY.SPGlaY, spontaneously hypertensive stroke-prone rats (SHRSP), and SP.WKYGlaY (n>5 per group). All relative gene expression data are expressed relative to the WKY group and are presented as mean±SEM. *P<0.05, **P<0.01, ***P<0.005 compared with WKY; #P<0.05, ##P<0.01 compared with SHRSP.

Article Snippet: Taqman predesigned assays (Applied Biosystems) for ACE (Rn00561094_m1), ACE2 (Rn01416293_m1), angiotensin II receptor type 1a (Rn02758772_s1), angiotensin II receptor type 2 (AT2R, Rn00560677_s1), and Mas receptor (MasR, Rn00562673_s1) were multiplexed with Actb (4352340E-labeled VIC).

Techniques: Gene Expression

Figure 3. Plasma angiotensin II (Ang II) and Ang (1-7) concentration, angiotensin-converting enzyme (ACE), and ACE2 activity in Wistar Kyoto (WKY), WKY.SPGlaY, spontaneously hypertensive stroke-prone rats (SHRSP), and SP.WKYGlaY (n≥5/ strain). Data are presented as mean±SEM with *P<0.05, **P<0.01 compared with WKY; #P<0.05, ##P<0.01 compared with SHRSP.

Journal: Hypertension

Article Title: Origin of the Y Chromosome Influences Intrarenal Vascular Responsiveness to Angiotensin I and Angiotensin (1-7) in Stroke-Prone Spontaneously Hypertensive Rats

doi: 10.1161/hypertensionaha.114.03756

Figure Lengend Snippet: Figure 3. Plasma angiotensin II (Ang II) and Ang (1-7) concentration, angiotensin-converting enzyme (ACE), and ACE2 activity in Wistar Kyoto (WKY), WKY.SPGlaY, spontaneously hypertensive stroke-prone rats (SHRSP), and SP.WKYGlaY (n≥5/ strain). Data are presented as mean±SEM with *P<0.05, **P<0.01 compared with WKY; #P<0.05, ##P<0.01 compared with SHRSP.

Article Snippet: Taqman predesigned assays (Applied Biosystems) for ACE (Rn00561094_m1), ACE2 (Rn01416293_m1), angiotensin II receptor type 1a (Rn02758772_s1), angiotensin II receptor type 2 (AT2R, Rn00560677_s1), and Mas receptor (MasR, Rn00562673_s1) were multiplexed with Actb (4352340E-labeled VIC).

Techniques: Clinical Proteomics, Concentration Assay, Activity Assay

mRNA target selection from the 30 probes included in the prognostically relevant signature and TaqMan® gene expression assay description.

Journal: PLoS ONE

Article Title: Identification and Validation of a Multigene Predictor of Recurrence in Primary Laryngeal Cancer

doi: 10.1371/journal.pone.0070429

Figure Lengend Snippet: mRNA target selection from the 30 probes included in the prognostically relevant signature and TaqMan® gene expression assay description.

Article Snippet: 1 , 219962_at , Angiotensin I converting enzyme (peptidyl-dipeptidase A) 2 , NM_021804 , ACE2 , 59272 , , 0,0660013 , 1,06 , yes , , NM_021804.2 , Hs01085334_m1 , 83 , 18–19 , Accepted.

Techniques: Selection, Gene Expression, Membrane, Binding Assay, Sequencing

30-probe set model annotations.

Journal: PLoS ONE

Article Title: Identification and Validation of a Multigene Predictor of Recurrence in Primary Laryngeal Cancer

doi: 10.1371/journal.pone.0070429

Figure Lengend Snippet: 30-probe set model annotations.

Article Snippet: 1 , 219962_at , Angiotensin I converting enzyme (peptidyl-dipeptidase A) 2 , NM_021804 , ACE2 , 59272 , , 0,0660013 , 1,06 , yes , , NM_021804.2 , Hs01085334_m1 , 83 , 18–19 , Accepted.

Techniques: Binding Assay, Membrane, Sequencing

Individual and profiled gene expression effects on DFS.

Journal: PLoS ONE

Article Title: Identification and Validation of a Multigene Predictor of Recurrence in Primary Laryngeal Cancer

doi: 10.1371/journal.pone.0070429

Figure Lengend Snippet: Individual and profiled gene expression effects on DFS.

Article Snippet: 1 , 219962_at , Angiotensin I converting enzyme (peptidyl-dipeptidase A) 2 , NM_021804 , ACE2 , 59272 , , 0,0660013 , 1,06 , yes , , NM_021804.2 , Hs01085334_m1 , 83 , 18–19 , Accepted.

Techniques: Gene Expression

Construction of HEK293T cells line that continuously expressing ACE2-GFP. HEK293Tcells were transfected with pCMV-ACE2-GFPSark tag plasmid and selected with hygromycin B to generate cell lines expressingACE2-GFP, cells were passaged 10 times to ensure stable expression. a Confocal imaging of HEK293T cell line with stably expressed ACE2-GFP. b Cell lysates were analyzed by Western blot with antibodies specific for ACE2 to confirm the expression of ACE2-GFP in HEK293T cells

Journal: Biological Procedures Online

Article Title: Investigation of Interaction between the Spike Protein of SARS-CoV-2 and ACE2-Expressing Cells Using an In Vitro Cell Capturing System

doi: 10.1186/s12575-021-00153-9

Figure Lengend Snippet: Construction of HEK293T cells line that continuously expressing ACE2-GFP. HEK293Tcells were transfected with pCMV-ACE2-GFPSark tag plasmid and selected with hygromycin B to generate cell lines expressingACE2-GFP, cells were passaged 10 times to ensure stable expression. a Confocal imaging of HEK293T cell line with stably expressed ACE2-GFP. b Cell lysates were analyzed by Western blot with antibodies specific for ACE2 to confirm the expression of ACE2-GFP in HEK293T cells

Article Snippet: HEK293T/ACE2 stable cell line was generated by transducing HEK293T cells with pCMV-Flag-ACE2,(Sino Biological, HG10108-NF), and 1 × 106 cells were lysed in lysis buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 2 mM EDTA, 1% NP-40, 1% sodium deoxycholate), supplemented with protease inhibitors.

Techniques: Expressing, Transfection, Plasmid Preparation, Imaging, Stable Transfection, Western Blot

Establishment of the in vitro cell capturing system using immobilized spikeS1 protein. a - e , Optimization of the dose of immobilized spike S1 protein for capturing HEK293T/ACE2-GFP cells. Different amounts of S1 protein were coated on the 96-well microplate to capture the HEK293T/ACE2-GFP cells. Representative micrographs of captured cells are shown for 0 μg (A),0.125 μg(B),0.25 μg (C),0.5 μg(D),and 1.0 μg(E). F Quantification of captured cells using CCK8 test, data are presented as mean ± SD

Journal: Biological Procedures Online

Article Title: Investigation of Interaction between the Spike Protein of SARS-CoV-2 and ACE2-Expressing Cells Using an In Vitro Cell Capturing System

doi: 10.1186/s12575-021-00153-9

Figure Lengend Snippet: Establishment of the in vitro cell capturing system using immobilized spikeS1 protein. a - e , Optimization of the dose of immobilized spike S1 protein for capturing HEK293T/ACE2-GFP cells. Different amounts of S1 protein were coated on the 96-well microplate to capture the HEK293T/ACE2-GFP cells. Representative micrographs of captured cells are shown for 0 μg (A),0.125 μg(B),0.25 μg (C),0.5 μg(D),and 1.0 μg(E). F Quantification of captured cells using CCK8 test, data are presented as mean ± SD

Article Snippet: HEK293T/ACE2 stable cell line was generated by transducing HEK293T cells with pCMV-Flag-ACE2,(Sino Biological, HG10108-NF), and 1 × 106 cells were lysed in lysis buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 2 mM EDTA, 1% NP-40, 1% sodium deoxycholate), supplemented with protease inhibitors.

Techniques: In Vitro

Competitive curve of cell capturing inhibition by RBD domain of Spike protein. 0.5 μg Spike S1 protein was coated on the microplate to capture the HEK293T/ACE2-GFP cells in presence of different concentrations of spike RBD protein. Amount of captured cells were determined by CCK8 test and expressed as relative value by setting the capability to capture cells of the non-RBD group as 100%. Data were plotted with a four-parameter logistic (4PL) regression curve fit of relative cell number (y-axis) versus the competitor concentration (x-axis)

Journal: Biological Procedures Online

Article Title: Investigation of Interaction between the Spike Protein of SARS-CoV-2 and ACE2-Expressing Cells Using an In Vitro Cell Capturing System

doi: 10.1186/s12575-021-00153-9

Figure Lengend Snippet: Competitive curve of cell capturing inhibition by RBD domain of Spike protein. 0.5 μg Spike S1 protein was coated on the microplate to capture the HEK293T/ACE2-GFP cells in presence of different concentrations of spike RBD protein. Amount of captured cells were determined by CCK8 test and expressed as relative value by setting the capability to capture cells of the non-RBD group as 100%. Data were plotted with a four-parameter logistic (4PL) regression curve fit of relative cell number (y-axis) versus the competitor concentration (x-axis)

Article Snippet: HEK293T/ACE2 stable cell line was generated by transducing HEK293T cells with pCMV-Flag-ACE2,(Sino Biological, HG10108-NF), and 1 × 106 cells were lysed in lysis buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 2 mM EDTA, 1% NP-40, 1% sodium deoxycholate), supplemented with protease inhibitors.

Techniques: Inhibition, Concentration Assay

Beas-2B cell migration was decreased by the treatment with SARS-CoV-2 (2019-nCoV) spike protein. (A, B) Beas-2B cells were transfected with pCMV control or pCMV3-ACE2-Flag plasmids DNA for 24 h. Cells were treated with 25 ng/mL SARS-CoV-2 (2019-nCoV) spike protein in the presence or absence of FBS for another 24 h. The cells were fixed and stained with antibody against Arl13b and DAPI. The ciliated cells were counted (n>500 cells) (A). Cell lysates were prepared and the protein expression of ACE2-Flag was detected by western blotting using antibody against Flag (B). (C-E) Beas-2B cells were plated on 35-mm 2 dishes and incubated for 24 h. A confluent monolayer of Beas-2B cells was then scratched with a sterile pipet tip. Then, cells were treated with 25 ng/mL spike protein and incubated for 24 h. Migration of cells into the space left by the scratch was photographed using a phase-contrast microscope at 200× magnification (C). Percentage of cell migration was quantified by subtracting the empty area remained at each time point from that at the initiation using NIH image analysis software (version 1.62; National Institutes of Health), and compared to that of the 0-h time point. Percentage of cell migration was presented as bar graph (D). Changes in single cell path were analyzed by tracking program, ImageJ plugin MTrackJ (Version 1.5.0). Movement track was presented in coordinates for each cell on a polar grid with the normalization of start points to the origin (E). Data in bar graphs represented as means ± SD. # p <0.05, significantly different from pCMV-transfected and spike protein-untreated group incubated with or without FBS & p <0.05, significantly different from pCMV3-ACE2-transfected and spike protein-untreated group incubated with or without FBS (A, D). ** p <0.01, significantly different from spike protein-untreated group (D).

Journal: Biomolecules & Therapeutics

Article Title: Primary Cilium by Polyinosinic:Polycytidylic Acid Regulates the Regenerative Migration of Beas-2B Bronchial Epithelial Cells

doi: 10.4062/biomolther.2022.009

Figure Lengend Snippet: Beas-2B cell migration was decreased by the treatment with SARS-CoV-2 (2019-nCoV) spike protein. (A, B) Beas-2B cells were transfected with pCMV control or pCMV3-ACE2-Flag plasmids DNA for 24 h. Cells were treated with 25 ng/mL SARS-CoV-2 (2019-nCoV) spike protein in the presence or absence of FBS for another 24 h. The cells were fixed and stained with antibody against Arl13b and DAPI. The ciliated cells were counted (n>500 cells) (A). Cell lysates were prepared and the protein expression of ACE2-Flag was detected by western blotting using antibody against Flag (B). (C-E) Beas-2B cells were plated on 35-mm 2 dishes and incubated for 24 h. A confluent monolayer of Beas-2B cells was then scratched with a sterile pipet tip. Then, cells were treated with 25 ng/mL spike protein and incubated for 24 h. Migration of cells into the space left by the scratch was photographed using a phase-contrast microscope at 200× magnification (C). Percentage of cell migration was quantified by subtracting the empty area remained at each time point from that at the initiation using NIH image analysis software (version 1.62; National Institutes of Health), and compared to that of the 0-h time point. Percentage of cell migration was presented as bar graph (D). Changes in single cell path were analyzed by tracking program, ImageJ plugin MTrackJ (Version 1.5.0). Movement track was presented in coordinates for each cell on a polar grid with the normalization of start points to the origin (E). Data in bar graphs represented as means ± SD. # p <0.05, significantly different from pCMV-transfected and spike protein-untreated group incubated with or without FBS & p <0.05, significantly different from pCMV3-ACE2-transfected and spike protein-untreated group incubated with or without FBS (A, D). ** p <0.01, significantly different from spike protein-untreated group (D).

Article Snippet: Human pCMV3-ACE2-Flag plasmid (HG10108-CF) and SARS-CoV-2 (2019-nCoV) spike protein (40592-V08B-B) were purchased from Sino Biological Inc (Wayne, PA, USA).

Techniques: Migration, Transfection, Staining, Expressing, Western Blot, Incubation, Microscopy, Software

BHK cells stably expressing GFP and hACE2 were incubated with His-tagged MERS-CoV RBD, SARS-CoV-2 WT RBD, Alpha RBD, Beta RBD, Gamma RBD, Mink-Y453F RBD, Mink-N501T RBD, and Mink-F486L RBD, respectively. APC anti-His antibodies were used to detect the His-tagged protein binding to the cells. Representative results from three experiments are shown. The mean ± SD percentages of RBD-binding cells in the three experiments are shown in the right lower bar chart. Statistical significance was analyzed using one-way ANOVA with a Tukey’s multiple comparison test for multiple groups.

Journal: Nature Communications

Article Title: Molecular insights into receptor binding of recent emerging SARS-CoV-2 variants

doi: 10.1038/s41467-021-26401-w

Figure Lengend Snippet: BHK cells stably expressing GFP and hACE2 were incubated with His-tagged MERS-CoV RBD, SARS-CoV-2 WT RBD, Alpha RBD, Beta RBD, Gamma RBD, Mink-Y453F RBD, Mink-N501T RBD, and Mink-F486L RBD, respectively. APC anti-His antibodies were used to detect the His-tagged protein binding to the cells. Representative results from three experiments are shown. The mean ± SD percentages of RBD-binding cells in the three experiments are shown in the right lower bar chart. Statistical significance was analyzed using one-way ANOVA with a Tukey’s multiple comparison test for multiple groups.

Article Snippet: Briefly, lentiviruses were packaged in HEK293T cells co-transfected with pLV-C-GFPSpark-hACE2 (Sino Biological) and helper plasmid pLP1, pLP2, and pLP/VSV-G (Invitrogen) at a ratio of 20:20:13:5 using Lipofectamine 2000.

Techniques: Stable Transfection, Expressing, Incubation, Protein Binding, Binding Assay

a – k Mouse Fc (mFc)-fused hACE2 or miACE2 in the supernatant was captured in the CM5 chip via its interaction with the pre-immobilized anti-mFc antibody. Various concentrations of SARS-CoV-2 WT RBD ( a ), Alpha RBD ( b ), Beta RBD ( c ), Gamma RBD ( d ), Mink-Y453F RBD ( e ), Mink-N501T RBD ( f ), and Mink-F486L RBD ( g ) protein were used to evaluate their binding affinity for hACE2. Serially diluted WT RBD ( h ), Mink-Y453F RBD ( i ), Mink-N501T RBD ( j ), and Mink-F486L RBD ( k ) protein were measured the binding to miACE2. K D , ka, and kd values are all recorded and the representative results from three experiments are shown. The data are presented as the mean ± SEM of three independent replicates ( n = 3).

Journal: Nature Communications

Article Title: Molecular insights into receptor binding of recent emerging SARS-CoV-2 variants

doi: 10.1038/s41467-021-26401-w

Figure Lengend Snippet: a – k Mouse Fc (mFc)-fused hACE2 or miACE2 in the supernatant was captured in the CM5 chip via its interaction with the pre-immobilized anti-mFc antibody. Various concentrations of SARS-CoV-2 WT RBD ( a ), Alpha RBD ( b ), Beta RBD ( c ), Gamma RBD ( d ), Mink-Y453F RBD ( e ), Mink-N501T RBD ( f ), and Mink-F486L RBD ( g ) protein were used to evaluate their binding affinity for hACE2. Serially diluted WT RBD ( h ), Mink-Y453F RBD ( i ), Mink-N501T RBD ( j ), and Mink-F486L RBD ( k ) protein were measured the binding to miACE2. K D , ka, and kd values are all recorded and the representative results from three experiments are shown. The data are presented as the mean ± SEM of three independent replicates ( n = 3).

Article Snippet: Briefly, lentiviruses were packaged in HEK293T cells co-transfected with pLV-C-GFPSpark-hACE2 (Sino Biological) and helper plasmid pLP1, pLP2, and pLP/VSV-G (Invitrogen) at a ratio of 20:20:13:5 using Lipofectamine 2000.

Techniques: Binding Assay

a – e The WT RBD-hACE2 structure (PDB: 6LZG) is shown in the center. Superimposition of WT RBD-hACE2 and each variant RBD-hACE2 (including Beta RBD-hACE2 ( a ), Gamma RBD-hACE2 ( b ), Alpha RBD-hACE2 ( c ), Mink-F486L RBD-hACE2 ( d ), Mink-Y453F RBD-hACE2 ( e )) are shown in each surrounding panel. In each structure the hACE2 is colored in light pink. SARS-CoV-2 WT RBD, Beta RBD, Gamma RBD, Alpha RBD, Mink-F486L RBD, and Mink-Y453F RBD are colored in gray, cyan, orange, yellow, green, and magenta, respectively. The key contact residues are shown as stick structures and labeled appropriately. The cation-π interaction, π-π stacking interaction, salt bridge, and hydrogen bonds are colored in magenta, blue, orange, and yellow, respectively. Hydrogen bond interactions were analyzed at a cutoff of 3.5 Å. f The detailed hydrogen bonds between the Mink-Y453F RBD and hACE2 are shown.

Journal: Nature Communications

Article Title: Molecular insights into receptor binding of recent emerging SARS-CoV-2 variants

doi: 10.1038/s41467-021-26401-w

Figure Lengend Snippet: a – e The WT RBD-hACE2 structure (PDB: 6LZG) is shown in the center. Superimposition of WT RBD-hACE2 and each variant RBD-hACE2 (including Beta RBD-hACE2 ( a ), Gamma RBD-hACE2 ( b ), Alpha RBD-hACE2 ( c ), Mink-F486L RBD-hACE2 ( d ), Mink-Y453F RBD-hACE2 ( e )) are shown in each surrounding panel. In each structure the hACE2 is colored in light pink. SARS-CoV-2 WT RBD, Beta RBD, Gamma RBD, Alpha RBD, Mink-F486L RBD, and Mink-Y453F RBD are colored in gray, cyan, orange, yellow, green, and magenta, respectively. The key contact residues are shown as stick structures and labeled appropriately. The cation-π interaction, π-π stacking interaction, salt bridge, and hydrogen bonds are colored in magenta, blue, orange, and yellow, respectively. Hydrogen bond interactions were analyzed at a cutoff of 3.5 Å. f The detailed hydrogen bonds between the Mink-Y453F RBD and hACE2 are shown.

Article Snippet: Briefly, lentiviruses were packaged in HEK293T cells co-transfected with pLV-C-GFPSpark-hACE2 (Sino Biological) and helper plasmid pLP1, pLP2, and pLP/VSV-G (Invitrogen) at a ratio of 20:20:13:5 using Lipofectamine 2000.

Techniques: Variant Assay, Labeling

The other does not include the transmembrane domain (pink) and lays nested between two motifs known to interact with the SARS-CoV receptor binding domain (blue). The measure of soluble ACE2 represents normalized expression of the pink to yellow target, or the proportion of non-transmembrane ACE2 to transmembrane ACE2 .

Journal: medRxiv

Article Title: The Contrasting Role of Nasopharyngeal Angiotensin Converting Enzyme 2 ( ACE2 ) Expression in SARS-CoV-2 Infection: A Cross-Sectional Study of People Tested for COVID-19 in British Columbia

doi: 10.1101/2020.11.23.20237206

Figure Lengend Snippet: The other does not include the transmembrane domain (pink) and lays nested between two motifs known to interact with the SARS-CoV receptor binding domain (blue). The measure of soluble ACE2 represents normalized expression of the pink to yellow target, or the proportion of non-transmembrane ACE2 to transmembrane ACE2 .

Article Snippet: Commercially available primer probe sets were used to amplify the host gene targets TMPRSS2 (Hs00237175_m1) (ThermoFisher) and ACE2 (Hs01085333_m1, HS01085340_m1) by multiplex qRT-PCR with a glyceraldehyde 3-phosphate dehydrogenase ( GAPDH ) control (Hs02758991_g1) (ThermoFisher).

Techniques: Binding Assay, Expressing

Boxplots of transmembrane ACE2 expression by nine-year age categories in COVID-19-negative participants between the ages of 19 and 98 (n=198); boxes represent the Q1-Q3 interquartile range, whiskers represent 1.5x the Q1 or Q3 and horizontal lines the median transmembrane ACE2 expression by age category. Nine-year categories were selected to optimize the distribution of observations between groups. Participants who tested negative younger than 19 or older than 98 were excluded based on n<10 observations per age group. No difference was detected in mean transmembrane ACE2 expression among age categories (ANOVA, P=0·092).

Journal: medRxiv

Article Title: The Contrasting Role of Nasopharyngeal Angiotensin Converting Enzyme 2 ( ACE2 ) Expression in SARS-CoV-2 Infection: A Cross-Sectional Study of People Tested for COVID-19 in British Columbia

doi: 10.1101/2020.11.23.20237206

Figure Lengend Snippet: Boxplots of transmembrane ACE2 expression by nine-year age categories in COVID-19-negative participants between the ages of 19 and 98 (n=198); boxes represent the Q1-Q3 interquartile range, whiskers represent 1.5x the Q1 or Q3 and horizontal lines the median transmembrane ACE2 expression by age category. Nine-year categories were selected to optimize the distribution of observations between groups. Participants who tested negative younger than 19 or older than 98 were excluded based on n<10 observations per age group. No difference was detected in mean transmembrane ACE2 expression among age categories (ANOVA, P=0·092).

Article Snippet: Commercially available primer probe sets were used to amplify the host gene targets TMPRSS2 (Hs00237175_m1) (ThermoFisher) and ACE2 (Hs01085333_m1, HS01085340_m1) by multiplex qRT-PCR with a glyceraldehyde 3-phosphate dehydrogenase ( GAPDH ) control (Hs02758991_g1) (ThermoFisher).

Techniques: Expressing

Gene expression is portrayed in kernel density plots stratified by COVID-19 test result. Probability densities of relative host gene expression are shown by positive (red, transmembrane ACE2 ; blue, soluble ACE2 ; yellow, TMPRSS2 ) and negative COVID-19 test results (grey). Levene’s test was used to detect non-equal variance in gene expression for all host targets between COVID-19-negative and -positive participants. A two-tailed, t-test was used to examine mean difference in host gene expression by COVID-19 test result assuming unequal variance: transmembrane ACE2 (P=1·2e-4), soluble ACE2 (P<0·0001) and TMPRSS2 (P<0·0001).

Journal: medRxiv

Article Title: The Contrasting Role of Nasopharyngeal Angiotensin Converting Enzyme 2 ( ACE2 ) Expression in SARS-CoV-2 Infection: A Cross-Sectional Study of People Tested for COVID-19 in British Columbia

doi: 10.1101/2020.11.23.20237206

Figure Lengend Snippet: Gene expression is portrayed in kernel density plots stratified by COVID-19 test result. Probability densities of relative host gene expression are shown by positive (red, transmembrane ACE2 ; blue, soluble ACE2 ; yellow, TMPRSS2 ) and negative COVID-19 test results (grey). Levene’s test was used to detect non-equal variance in gene expression for all host targets between COVID-19-negative and -positive participants. A two-tailed, t-test was used to examine mean difference in host gene expression by COVID-19 test result assuming unequal variance: transmembrane ACE2 (P=1·2e-4), soluble ACE2 (P<0·0001) and TMPRSS2 (P<0·0001).

Article Snippet: Commercially available primer probe sets were used to amplify the host gene targets TMPRSS2 (Hs00237175_m1) (ThermoFisher) and ACE2 (Hs01085333_m1, HS01085340_m1) by multiplex qRT-PCR with a glyceraldehyde 3-phosphate dehydrogenase ( GAPDH ) control (Hs02758991_g1) (ThermoFisher).

Techniques: Gene Expression, Two Tailed Test

The level of transmembrane ACE2 (red), soluble ACE2 (blue) and TMPRSS2 (black) expression in positive cases of COVID-19 and its relationship with SARS-CoV-2 RNA is shown in Log 10 genome equivalents per millilitre (GE/mL). Host gene expression was measured by quantitative real-time PCR and normalized to expression of the housekeeping gene GAPDH by the (2 -ΔΔCt ) method. Soluble ACE2 was defined as absolute relative expression between the Hs01085333_m1 (ThermoFisher) and HS01085340_m1 (ThermoFisher) gene targets . Simple linear regression was used estimate correlations, the reported P values are for the B eta-coefficients of host gene expression.

Journal: medRxiv

Article Title: The Contrasting Role of Nasopharyngeal Angiotensin Converting Enzyme 2 ( ACE2 ) Expression in SARS-CoV-2 Infection: A Cross-Sectional Study of People Tested for COVID-19 in British Columbia

doi: 10.1101/2020.11.23.20237206

Figure Lengend Snippet: The level of transmembrane ACE2 (red), soluble ACE2 (blue) and TMPRSS2 (black) expression in positive cases of COVID-19 and its relationship with SARS-CoV-2 RNA is shown in Log 10 genome equivalents per millilitre (GE/mL). Host gene expression was measured by quantitative real-time PCR and normalized to expression of the housekeeping gene GAPDH by the (2 -ΔΔCt ) method. Soluble ACE2 was defined as absolute relative expression between the Hs01085333_m1 (ThermoFisher) and HS01085340_m1 (ThermoFisher) gene targets . Simple linear regression was used estimate correlations, the reported P values are for the B eta-coefficients of host gene expression.

Article Snippet: Commercially available primer probe sets were used to amplify the host gene targets TMPRSS2 (Hs00237175_m1) (ThermoFisher) and ACE2 (Hs01085333_m1, HS01085340_m1) by multiplex qRT-PCR with a glyceraldehyde 3-phosphate dehydrogenase ( GAPDH ) control (Hs02758991_g1) (ThermoFisher).

Techniques: Expressing, Gene Expression, Real-time Polymerase Chain Reaction

Soluble ACE2 expression was categorized into low, mean and high levels to demonstrate the relationship. The low category (grey) represents soluble ACE2 expression two standard deviations below the mean expression. The mean category (orange) codes for the mean soluble ACE2 expression, zero standard deviations. The high category (blue) indicates soluble ACE2 expression two standard deviations above mean expression. Shaded areas represent 95% confidence intervals, solid lines represent B coefficients from multiple linear regression.

Journal: medRxiv

Article Title: The Contrasting Role of Nasopharyngeal Angiotensin Converting Enzyme 2 ( ACE2 ) Expression in SARS-CoV-2 Infection: A Cross-Sectional Study of People Tested for COVID-19 in British Columbia

doi: 10.1101/2020.11.23.20237206

Figure Lengend Snippet: Soluble ACE2 expression was categorized into low, mean and high levels to demonstrate the relationship. The low category (grey) represents soluble ACE2 expression two standard deviations below the mean expression. The mean category (orange) codes for the mean soluble ACE2 expression, zero standard deviations. The high category (blue) indicates soluble ACE2 expression two standard deviations above mean expression. Shaded areas represent 95% confidence intervals, solid lines represent B coefficients from multiple linear regression.

Article Snippet: Commercially available primer probe sets were used to amplify the host gene targets TMPRSS2 (Hs00237175_m1) (ThermoFisher) and ACE2 (Hs01085333_m1, HS01085340_m1) by multiplex qRT-PCR with a glyceraldehyde 3-phosphate dehydrogenase ( GAPDH ) control (Hs02758991_g1) (ThermoFisher).

Techniques: Expressing