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R&D Systems human ace2 af 647 conjugated antibody

Figure 1. <t>ACE2,</t> HS, and SDC expression proﬁles of 293T and 293T-ACE2 cells. ACE2, HS, and SDC expression in 293T and 293T-ACE2 cells were assessed using ﬂuorescently labeled speciﬁc antibodies with imaging ﬂow cytometry. (A,B) ACE2 and HS expression proﬁle of 293T and 293T-ACE2 cells. The representative ﬂow cytometry histograms and cellular images show the ACE2 and HS expression of 293T and 293T-ACE2 cells treated with the ﬂuorescent ACE2 and HS antibodies. Scale bar = 20 µm. (C) SDC expression proﬁle of 293T and 293T-ACE2 cells. The representative ﬂow cytometry his- tograms and ﬂuorescent cellular images show the SDC expression of 293T cells and 293T-ACE2 cells treated with the APC-labeled respective SDC antibodies. Scale bar = 20 µm. (D–F) Detected ACE2, HS, and SDC expression values were normalized to those of 293T cells as standards. The bars represent the mean ± SEM of three independent experiments (data are represented as dots). Statistical signiﬁcance vs. standards was assessed with ANOVA. * p < 0.05; *** p < 0.001.

Human Ace2 Af 647 Conjugated Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human ace2 af 647 conjugated antibody - by Bioz Stars, 2026-09

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1) Product Images from "Exploring the Syndecan-Mediated Cellular Internalization of the SARS-CoV-2 Omicron Variant."

Article Title: Exploring the Syndecan-Mediated Cellular Internalization of the SARS-CoV-2 Omicron Variant.

Journal: International journal of molecular sciences

doi: 10.3390/ijms241814140

Figure 1. ACE2, HS, and SDC expression proﬁles of 293T and 293T-ACE2 cells. ACE2, HS, and SDC expression in 293T and 293T-ACE2 cells were assessed using ﬂuorescently labeled speciﬁc antibodies with imaging ﬂow cytometry. (A,B) ACE2 and HS expression proﬁle of 293T and 293T-ACE2 cells. The representative ﬂow cytometry histograms and cellular images show the ACE2 and HS expression of 293T and 293T-ACE2 cells treated with the ﬂuorescent ACE2 and HS antibodies. Scale bar = 20 µm. (C) SDC expression proﬁle of 293T and 293T-ACE2 cells. The representative ﬂow cytometry his- tograms and ﬂuorescent cellular images show the SDC expression of 293T cells and 293T-ACE2 cells treated with the APC-labeled respective SDC antibodies. Scale bar = 20 µm. (D–F) Detected ACE2, HS, and SDC expression values were normalized to those of 293T cells as standards. The bars represent the mean ± SEM of three independent experiments (data are represented as dots). Statistical signiﬁcance vs. standards was assessed with ANOVA. * p < 0.05; *** p < 0.001.

Figure Legend Snippet: Figure 1. ACE2, HS, and SDC expression proﬁles of 293T and 293T-ACE2 cells. ACE2, HS, and SDC expression in 293T and 293T-ACE2 cells were assessed using ﬂuorescently labeled speciﬁc antibodies with imaging ﬂow cytometry. (A,B) ACE2 and HS expression proﬁle of 293T and 293T-ACE2 cells. The representative ﬂow cytometry histograms and cellular images show the ACE2 and HS expression of 293T and 293T-ACE2 cells treated with the ﬂuorescent ACE2 and HS antibodies. Scale bar = 20 µm. (C) SDC expression proﬁle of 293T and 293T-ACE2 cells. The representative ﬂow cytometry his- tograms and ﬂuorescent cellular images show the SDC expression of 293T cells and 293T-ACE2 cells treated with the APC-labeled respective SDC antibodies. Scale bar = 20 µm. (D–F) Detected ACE2, HS, and SDC expression values were normalized to those of 293T cells as standards. The bars represent the mean ± SEM of three independent experiments (data are represented as dots). Statistical signiﬁcance vs. standards was assessed with ANOVA. * p < 0.05; *** p < 0.001.

Techniques Used: Expressing, Labeling, Imaging, Cytometry

Figure 2. Cellular uptake of WT SCV2, Delta, and Omicron variants into 293T- and ACE2- overexpressing 293T-ACE2 cells. The cells were exposed to 1 MOI of heat-inactivated WT SCV2, Delta, and Omicron variants for 4 h at 37 ◦C. After incubation, the cells were washed, trypsinized, ﬁxed, permeabilized, and treated with a primary SARS-CoV-2 spike (1000–1200 aa), followed by a ﬂuorescently labeled (AF 633) secondary antibody. Cellular uptake was then analyzed with imaging ﬂow cytometry. (A–D) Representative ﬂow cytometry histograms and brightﬁeld (BF) and ﬂuorescent cellular images showing the intracellular ﬂuorescence of the virus-exposed 293T and 293T-ACE2 cells. Scale bar = 20 µm. (E) Detected ﬂuorescence intensities were normalized to WT SCV2-treated 293T cells as standards. The bars represent the mean + SEM of ﬁve independent experiments. Experimental data are presented as dots. Statistical signiﬁcance was assessed with ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001; ns: not signiﬁcant.

Figure Legend Snippet: Figure 2. Cellular uptake of WT SCV2, Delta, and Omicron variants into 293T- and ACE2- overexpressing 293T-ACE2 cells. The cells were exposed to 1 MOI of heat-inactivated WT SCV2, Delta, and Omicron variants for 4 h at 37 ◦C. After incubation, the cells were washed, trypsinized, ﬁxed, permeabilized, and treated with a primary SARS-CoV-2 spike (1000–1200 aa), followed by a ﬂuorescently labeled (AF 633) secondary antibody. Cellular uptake was then analyzed with imaging ﬂow cytometry. (A–D) Representative ﬂow cytometry histograms and brightﬁeld (BF) and ﬂuorescent cellular images showing the intracellular ﬂuorescence of the virus-exposed 293T and 293T-ACE2 cells. Scale bar = 20 µm. (E) Detected ﬂuorescence intensities were normalized to WT SCV2-treated 293T cells as standards. The bars represent the mean + SEM of ﬁve independent experiments. Experimental data are presented as dots. Statistical signiﬁcance was assessed with ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001; ns: not signiﬁcant.

Techniques Used: Incubation, Labeling, Imaging, Cytometry, Virus

Expressing:

Article Title: Exploring the Syndecan-Mediated Cellular Internalization of the SARS-CoV-2 Omicron Variant.
Article Snippet: .. ACE2 expression was measured with human ACE2 AF 647-conjugated antibody (RnD Systems, Minneapolis, MN, USA, cat. no. FAB9332R) and respective isotype control (mouse IgG2A AF 647-conjugated isotype control, RnD Systems, cat. no. IC003R), according to the manufacturer’s protocol. .. SDC4 knockdown in Calu-3 cells was performed as described previously, using a lentiviral vector system specific to human SDC4 shRNA (cat. nos. sc-41400-SH an sc36588), according to the manufacturer’s protocol (Santa Cruz Biotechnology, Inc., Dallas, TX, USA) [37].

Article Title: Exploring the Syndecan-Mediated Cellular Internalization of the SARS-CoV-2 Omicron Variant
Article Snippet: .. ACE2 expression was measured with human ACE2 AF 647-conjugated antibody (RnD Systems, Minneapolis, MN, USA, cat. no. FAB9332R) and respective isotype control (mouse IgG2A AF 647-conjugated isotype control, RnD Systems, cat. no. IC003R), according to the manufacturer’s protocol. .. SDC4 knockdown in Calu-3 cells was performed as described previously, using a lentiviral vector system specific to human SDC4 shRNA (cat. nos. sc-41400-SH an sc-36588), according to the manufacturer’s protocol (Santa Cruz Biotechnology, Inc., Dallas, TX, USA) [ ].

Article Title: Syndecan-4 Is a Key Facilitator of the SARS-CoV-2 Delta Variant’s Superior Transmission
Article Snippet: .. ACE2 expression was measured with human ACE2 AF 647-conjugated antibody (RnD Systems, Minneapolis, MN, USA, cat. no. FAB9332R) and respective isotype control (mouse IgG2A AF 647-conjugated isotype control, RnD Systems, cat. no. IC003R), according to the manufacturer’s protocol. .. SDC4 knockdown in Calu-3 cells was performed using a lentiviral vector system specific to human SDC4 shRNA (cat. no. sc-41400-SH, sc-36588), according to the manufacturer’s protocol (Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

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Journal: International journal of molecular sciences

Article Title: Exploring the Syndecan-Mediated Cellular Internalization of the SARS-CoV-2 Omicron Variant.

doi: 10.3390/ijms241814140

Figure Lengend Snippet: Figure 1. ACE2, HS, and SDC expression proﬁles of 293T and 293T-ACE2 cells. ACE2, HS, and SDC expression in 293T and 293T-ACE2 cells were assessed using ﬂuorescently labeled speciﬁc antibodies with imaging ﬂow cytometry. (A,B) ACE2 and HS expression proﬁle of 293T and 293T-ACE2 cells. The representative ﬂow cytometry histograms and cellular images show the ACE2 and HS expression of 293T and 293T-ACE2 cells treated with the ﬂuorescent ACE2 and HS antibodies. Scale bar = 20 µm. (C) SDC expression proﬁle of 293T and 293T-ACE2 cells. The representative ﬂow cytometry his- tograms and ﬂuorescent cellular images show the SDC expression of 293T cells and 293T-ACE2 cells treated with the APC-labeled respective SDC antibodies. Scale bar = 20 µm. (D–F) Detected ACE2, HS, and SDC expression values were normalized to those of 293T cells as standards. The bars represent the mean ± SEM of three independent experiments (data are represented as dots). Statistical signiﬁcance vs. standards was assessed with ANOVA. * p < 0.05; *** p < 0.001.

Article Snippet: ACE2 expression was measured with human ACE2 AF 647-conjugated antibody (RnD Systems, Minneapolis, MN, USA, cat. no. FAB9332R) and respective isotype control (mouse IgG2A AF 647-conjugated isotype control, RnD Systems, cat. no. IC003R), according to the manufacturer’s protocol.

Techniques: Expressing, Labeling, Imaging, Cytometry

Journal: International journal of molecular sciences

Article Title: Exploring the Syndecan-Mediated Cellular Internalization of the SARS-CoV-2 Omicron Variant.

doi: 10.3390/ijms241814140

Figure Lengend Snippet: Figure 2. Cellular uptake of WT SCV2, Delta, and Omicron variants into 293T- and ACE2- overexpressing 293T-ACE2 cells. The cells were exposed to 1 MOI of heat-inactivated WT SCV2, Delta, and Omicron variants for 4 h at 37 ◦C. After incubation, the cells were washed, trypsinized, ﬁxed, permeabilized, and treated with a primary SARS-CoV-2 spike (1000–1200 aa), followed by a ﬂuorescently labeled (AF 633) secondary antibody. Cellular uptake was then analyzed with imaging ﬂow cytometry. (A–D) Representative ﬂow cytometry histograms and brightﬁeld (BF) and ﬂuorescent cellular images showing the intracellular ﬂuorescence of the virus-exposed 293T and 293T-ACE2 cells. Scale bar = 20 µm. (E) Detected ﬂuorescence intensities were normalized to WT SCV2-treated 293T cells as standards. The bars represent the mean + SEM of ﬁve independent experiments. Experimental data are presented as dots. Statistical signiﬁcance was assessed with ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001; ns: not signiﬁcant.

Techniques: Incubation, Labeling, Imaging, Cytometry, Virus

Summary of sero-negative, acute, and convalescent-phase of COVID-19 patients from which the plasma ACE2 + EVs and RBD-IgG levels were measured.

Journal: Nature Communications

Article Title: Circulating ACE2-expressing extracellular vesicles block broad strains of SARS-CoV-2

doi: 10.1038/s41467-021-27893-2

Figure Lengend Snippet: Summary of sero-negative, acute, and convalescent-phase of COVID-19 patients from which the plasma ACE2 + EVs and RBD-IgG levels were measured.

Article Snippet: Cells were blocked with mouse serum IgG (Sigma, 15381) for 10 min at room temperature and then incubated with specific antibodies; AF-647 mouse anti-human ACE2 (Clone # 535919) (R&D systems, FAB9332R), AF-488 mouse anti-human ACE2 (Clone # 171607) (R&D systems, FAB9333G) (0.4 μg/10 6 cells), AF-647 isotype control mouse IgG2b (Clone # 20102) (R&D systems, IC003R) or AF-488 isotype control mouse IgG2bAF488 (Clone # 20102) (R&D systems, IC003G) for 45 min on ice, followed by washing twice with 2% EV-free FBS/PBS.

Techniques: Clinical Proteomics, Sampling

a ACE2+ EVs detected in human plasma samples of sero-negative controls (light blue), acute phase (dark green), and convalescent COVID-19 patients (green). One-tail t test (* p = 0.038, ** p = 0.0061 and ** p = 0.0016). Data are presented as mean values ± SEM. b Representative microflow vesiclometry (MFV) plots with gated ACE2+ EVs from sero-negative, acute phase and convalescent COVID-19 patients. c MFV detection of circulating ACE2 + EVs with CD63 + EVs in human plasma of convalescent COVID-19 patient samples (CSB-029 and CSB-023) (green line). Blue line is isotype IgG-negative control. d Flow profiles of ACE2 expression in HEK and HeLa parental control cells (Con, light blue line, ACE2 − ) and with ACE2 overexpression (ACE2, green line). e NanoSight NTA analysis of the sizes of HEK-derived ACE2 − (ev1Con) and ACE2 + (ev1ACE2) and HeLa-derived ACE2 − (ev2Con) and ACE2 + (ev2ACE2). f Immunoblots of HEK and HeLa (ACE2 − and ACE2 + ) EVs and cell lysates for ACE2, TSG101, CD63, CD81, GRP94 and loading control of the membrane proteins upon Ponceau staining. RIPA buffer and Bradford protein assay were used for cells/EVs lysis and protein measurement, respectively ( N = 1 experiment). g Cryo-EM images of HEK-derived EVs, ACE2 − (evCon, left) and ACE2 + (evACE2, right), stained with ACE2 (top) and CD81 (bottom). Scale bars = 100 nm. h Quantified counts of Apogee MFV-based total extracellular vesicles (EVs) and ACE2 + EVs ( N = 2 experiments with n = 6 technical replicates for total EV particles and n = 3 technical replicates for ACE2 + counts). Control EVs are in light blue and ACE2 + EVs in green. Data are presented as mean values +/− SD. i Overlay flow profiles of ACE2 positivity within CD63 + (left column) and CD81+ (right column) EVs isolated from HEK-ACE2 (top row) and HeLa-ACE2 (bottom row) cells, respectively ( n = 3 technical replicates). Light blue line for Control EVs and green line for ACE2 + EVs.

Journal: Nature Communications

Article Title: Circulating ACE2-expressing extracellular vesicles block broad strains of SARS-CoV-2

doi: 10.1038/s41467-021-27893-2

Figure Lengend Snippet: a ACE2+ EVs detected in human plasma samples of sero-negative controls (light blue), acute phase (dark green), and convalescent COVID-19 patients (green). One-tail t test (* p = 0.038, ** p = 0.0061 and ** p = 0.0016). Data are presented as mean values ± SEM. b Representative microflow vesiclometry (MFV) plots with gated ACE2+ EVs from sero-negative, acute phase and convalescent COVID-19 patients. c MFV detection of circulating ACE2 + EVs with CD63 + EVs in human plasma of convalescent COVID-19 patient samples (CSB-029 and CSB-023) (green line). Blue line is isotype IgG-negative control. d Flow profiles of ACE2 expression in HEK and HeLa parental control cells (Con, light blue line, ACE2 − ) and with ACE2 overexpression (ACE2, green line). e NanoSight NTA analysis of the sizes of HEK-derived ACE2 − (ev1Con) and ACE2 + (ev1ACE2) and HeLa-derived ACE2 − (ev2Con) and ACE2 + (ev2ACE2). f Immunoblots of HEK and HeLa (ACE2 − and ACE2 + ) EVs and cell lysates for ACE2, TSG101, CD63, CD81, GRP94 and loading control of the membrane proteins upon Ponceau staining. RIPA buffer and Bradford protein assay were used for cells/EVs lysis and protein measurement, respectively ( N = 1 experiment). g Cryo-EM images of HEK-derived EVs, ACE2 − (evCon, left) and ACE2 + (evACE2, right), stained with ACE2 (top) and CD81 (bottom). Scale bars = 100 nm. h Quantified counts of Apogee MFV-based total extracellular vesicles (EVs) and ACE2 + EVs ( N = 2 experiments with n = 6 technical replicates for total EV particles and n = 3 technical replicates for ACE2 + counts). Control EVs are in light blue and ACE2 + EVs in green. Data are presented as mean values +/− SD. i Overlay flow profiles of ACE2 positivity within CD63 + (left column) and CD81+ (right column) EVs isolated from HEK-ACE2 (top row) and HeLa-ACE2 (bottom row) cells, respectively ( n = 3 technical replicates). Light blue line for Control EVs and green line for ACE2 + EVs.

Techniques: Clinical Proteomics, Negative Control, Expressing, Control, Over Expression, Derivative Assay, Western Blot, Membrane, Staining, Bradford Protein Assay, Lysis, Cryo-EM Sample Prep, Isolation

a Schematic depiction of the cell-based neutralization assay. b Representative flow profiles showing the percentage (fluorescence mean intensity) of RBD-AF647 binding (at 16 and 3.3 nmol/L) to ACE2 + HEK-293 cells, inhibited by rhACE2 and ACE2 + EVs (evACE2) isolated from HEK-293 and HeLa cells (HEK-EV1 and HeLa-EV2, respectively) whereas ACE2 − EVs (evCon) had no neutralization effects (no RBD in black, PBS in dark blue, rhACE2 in orange, evCon in light blue, and evACE2 in green). c IC 50 of rhACE2 (orange line) and ACE2 in the EVs from ACE2 + HEK (ev1ACE2) and HeLa (ev2ACE2) cells (green lines) on 16 nM RBD-host cell binding (%). GraphPad Prism 9.0.2 was used to calculate the IC 50 . N = 2 experiments with two technical replicates for each. Data are presented as mean values ± SD. d IC 50 of evACE2, ev1 from HEK and ev2 from HeLa cells (green lines), and rhACE2 (orange line) neutralizing infections by wild-type (WT) S + pseudotyped SARS-CoV-2. GraphPad Prism 9.0.2 was used to calculate the IC 50 . N = 2 experiments with two technical replicates for each. Data are presented as mean values ± SD. e IC 50 (nM) of ACE2 in ev1ACE2 (HEK) (green line) and rhACE2 (orange line) upon wild-type SARS-CoV-2 infection. GraphPad Prism 9.0.2 was used to calculate the IC 50 with three biological replicates. Data are presented as mean values ± SD. f Distinct effects of ACE2 + EVs (green lines) and ACE2 − control EVs (light blue line) on inhibiting Vero-6 cell death caused by SARS-CoV-2. N = 2 experiments with three biological replicates each. Data are presented as mean values ± SD. g The IC 50 of ev1ACE2 (HEK) neutralizing infections by pseudotyped SARS-CoV-2 expressing WT (black), B.1.1.7 (α) variant (red), B1.351 (β) variant (dark blue) and B.1.617.2 (δ) (light green) S protein. GraphPad Prism 9.0.2 was used to calculate the IC 50 . N = 2 experiments with two technical replicates each. Data are presented as mean values ± SD. h Effects of ev1ACE2 (HEK) on protecting Vero-6 cell viability against infections of SARS-CoV-2 WT (black), B.1.1.7 (α) variant (red) and B1.351 (β) variant (dark blue) ( n = 3 biological replicates). Data are presented as mean values ± SD.

Journal: Nature Communications

Article Title: Circulating ACE2-expressing extracellular vesicles block broad strains of SARS-CoV-2

doi: 10.1038/s41467-021-27893-2

Figure Lengend Snippet: a Schematic depiction of the cell-based neutralization assay. b Representative flow profiles showing the percentage (fluorescence mean intensity) of RBD-AF647 binding (at 16 and 3.3 nmol/L) to ACE2 + HEK-293 cells, inhibited by rhACE2 and ACE2 + EVs (evACE2) isolated from HEK-293 and HeLa cells (HEK-EV1 and HeLa-EV2, respectively) whereas ACE2 − EVs (evCon) had no neutralization effects (no RBD in black, PBS in dark blue, rhACE2 in orange, evCon in light blue, and evACE2 in green). c IC 50 of rhACE2 (orange line) and ACE2 in the EVs from ACE2 + HEK (ev1ACE2) and HeLa (ev2ACE2) cells (green lines) on 16 nM RBD-host cell binding (%). GraphPad Prism 9.0.2 was used to calculate the IC 50 . N = 2 experiments with two technical replicates for each. Data are presented as mean values ± SD. d IC 50 of evACE2, ev1 from HEK and ev2 from HeLa cells (green lines), and rhACE2 (orange line) neutralizing infections by wild-type (WT) S + pseudotyped SARS-CoV-2. GraphPad Prism 9.0.2 was used to calculate the IC 50 . N = 2 experiments with two technical replicates for each. Data are presented as mean values ± SD. e IC 50 (nM) of ACE2 in ev1ACE2 (HEK) (green line) and rhACE2 (orange line) upon wild-type SARS-CoV-2 infection. GraphPad Prism 9.0.2 was used to calculate the IC 50 with three biological replicates. Data are presented as mean values ± SD. f Distinct effects of ACE2 + EVs (green lines) and ACE2 − control EVs (light blue line) on inhibiting Vero-6 cell death caused by SARS-CoV-2. N = 2 experiments with three biological replicates each. Data are presented as mean values ± SD. g The IC 50 of ev1ACE2 (HEK) neutralizing infections by pseudotyped SARS-CoV-2 expressing WT (black), B.1.1.7 (α) variant (red), B1.351 (β) variant (dark blue) and B.1.617.2 (δ) (light green) S protein. GraphPad Prism 9.0.2 was used to calculate the IC 50 . N = 2 experiments with two technical replicates each. Data are presented as mean values ± SD. h Effects of ev1ACE2 (HEK) on protecting Vero-6 cell viability against infections of SARS-CoV-2 WT (black), B.1.1.7 (α) variant (red) and B1.351 (β) variant (dark blue) ( n = 3 biological replicates). Data are presented as mean values ± SD.

Techniques: Neutralization, Fluorescence, Binding Assay, Isolation, Infection, Control, Expressing, Variant Assay

a Schematic depiction of plasma EV ultracentrifugation and RBD-bead based depletion. b Cryo-EM images of human EV pellets isolated from acute phase COVID-19 plasma (bar = 100 nm). c Immunoblots of plasma EV pellets (sero-negative and COVID-19 acute phase patients CBB-005 and -013) for ACE2 and loading control of protein staining with Ponceau). Laemmli buffer was used for lysis ( N = 1 experiment). d ACE2 + EV pellets from acute phase patients 007, 008, 009, 012, and 013 (CBB) ( n = 2 biological replicates each) blocked SARS-CoV-2 infection-induced death of Vero-6 cells whereas the sero-negative control ( n = 2 biological replicates) and CBB-005 (no detectable ACE2) ( n = 2 biological replicates) did not show neutralization effects. One-tail t test, **** p = 2.24E−08 shown as compared to sero-negative. e , f Levels of ACE2 + EV counts ( n = 3 biological replicates) in plasma EVs (green) and bead-depleted EVs (light blue). One-tail paired t test, * p = 0.011 and ** p = 0.0063 (data are presented as mean values ± SD) ( e ) and altered neutralization effects on RBD–host cell binding ( f ) of the COVID-19 plasma EV pellets prior to and after RBD-bead depletion (convalescent phase CSB-012 and -024; acute phase CBB-008, 009, and 013). One-tail paired t test **** p = 5.11E−05.

Journal: Nature Communications

Article Title: Circulating ACE2-expressing extracellular vesicles block broad strains of SARS-CoV-2

doi: 10.1038/s41467-021-27893-2

Figure Lengend Snippet: a Schematic depiction of plasma EV ultracentrifugation and RBD-bead based depletion. b Cryo-EM images of human EV pellets isolated from acute phase COVID-19 plasma (bar = 100 nm). c Immunoblots of plasma EV pellets (sero-negative and COVID-19 acute phase patients CBB-005 and -013) for ACE2 and loading control of protein staining with Ponceau). Laemmli buffer was used for lysis ( N = 1 experiment). d ACE2 + EV pellets from acute phase patients 007, 008, 009, 012, and 013 (CBB) ( n = 2 biological replicates each) blocked SARS-CoV-2 infection-induced death of Vero-6 cells whereas the sero-negative control ( n = 2 biological replicates) and CBB-005 (no detectable ACE2) ( n = 2 biological replicates) did not show neutralization effects. One-tail t test, **** p = 2.24E−08 shown as compared to sero-negative. e , f Levels of ACE2 + EV counts ( n = 3 biological replicates) in plasma EVs (green) and bead-depleted EVs (light blue). One-tail paired t test, * p = 0.011 and ** p = 0.0063 (data are presented as mean values ± SD) ( e ) and altered neutralization effects on RBD–host cell binding ( f ) of the COVID-19 plasma EV pellets prior to and after RBD-bead depletion (convalescent phase CSB-012 and -024; acute phase CBB-008, 009, and 013). One-tail paired t test **** p = 5.11E−05.

Techniques: Clinical Proteomics, Cryo-EM Sample Prep, Isolation, Western Blot, Control, Staining, Lysis, Infection, Negative Control, Neutralization, Binding Assay

a Probability of severe disease-free survival in B6.Cg-Tg(K18-ACE2)2Prlmn/J (K18-hACE2) mice receiving SARS-CoV-2 infection (10,000 pfu) and intranasal EVs (130 µg as measured on Nanodrop) per mouse (evCon in light blue and evACE2 in green). Log-rank (Mantel–Cox) and Gehan–Breslow–Wilcoxon tests **** p = 2.27E−07. b Viral loads in mouse lungs on day 5/6 after receiving SARS-CoV-2 infection and administration of evCon ( N = 5 mice) (light blue) or evACE2 ( N = 10 mice) (green). T -test–nonparametric-one tailed, * p = 0.013. Data are presented as mean values ± SD. c . Representative H&E images of mouse lung sections at day 5 or 6 post virus inoculation and EV treatment (evCon and evACE2) intranasally. d , e Acute and chronic inflammation scores ( d ), and alveolar hemorrhage and necrosis scores ( e ) in mouse lungs on day 5/6 after receiving evCon ( N = 5 mice) (light blue) or evACE2 ( N = 7 mice) (green). T -test–nonparametric-one tailed, ** p = 0.005, ** p = 0.003 and *** p = 0.0004. Data are presented as mean values ± SD.

Journal: Nature Communications

Article Title: Circulating ACE2-expressing extracellular vesicles block broad strains of SARS-CoV-2

doi: 10.1038/s41467-021-27893-2

Figure Lengend Snippet: a Probability of severe disease-free survival in B6.Cg-Tg(K18-ACE2)2Prlmn/J (K18-hACE2) mice receiving SARS-CoV-2 infection (10,000 pfu) and intranasal EVs (130 µg as measured on Nanodrop) per mouse (evCon in light blue and evACE2 in green). Log-rank (Mantel–Cox) and Gehan–Breslow–Wilcoxon tests **** p = 2.27E−07. b Viral loads in mouse lungs on day 5/6 after receiving SARS-CoV-2 infection and administration of evCon ( N = 5 mice) (light blue) or evACE2 ( N = 10 mice) (green). T -test–nonparametric-one tailed, * p = 0.013. Data are presented as mean values ± SD. c . Representative H&E images of mouse lung sections at day 5 or 6 post virus inoculation and EV treatment (evCon and evACE2) intranasally. d , e Acute and chronic inflammation scores ( d ), and alveolar hemorrhage and necrosis scores ( e ) in mouse lungs on day 5/6 after receiving evCon ( N = 5 mice) (light blue) or evACE2 ( N = 7 mice) (green). T -test–nonparametric-one tailed, ** p = 0.005, ** p = 0.003 and *** p = 0.0004. Data are presented as mean values ± SD.

Techniques: Infection, One-tailed Test, Virus

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human ace2 af 647 conjugated antibody (R&D Systems)

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1) Product Images from "Exploring the Syndecan-Mediated Cellular Internalization of the SARS-CoV-2 Omicron Variant."

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