hace2  (Bio-Techne corporation)


Bioz Verified Symbol Bio-Techne corporation is a verified supplier
Bioz Manufacturer Symbol Bio-Techne corporation manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96

    Structured Review

    Bio-Techne corporation hace2
    Hace2, supplied by Bio-Techne corporation, 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/result/hace2/product/Bio-Techne corporation
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hace2 - by Bioz Stars, 2024-05
    96/100 stars

    Images

    hace2  (Bio-Techne corporation)


    Bioz Verified Symbol Bio-Techne corporation is a verified supplier
    Bioz Manufacturer Symbol Bio-Techne corporation manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96

    Structured Review

    Bio-Techne corporation hace2
    Hace2, supplied by Bio-Techne corporation, 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/result/hace2/product/Bio-Techne corporation
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hace2 - by Bioz Stars, 2024-05
    96/100 stars

    Images

    hace2  (Bio-Techne corporation)


    Bioz Verified Symbol Bio-Techne corporation is a verified supplier
    Bioz Manufacturer Symbol Bio-Techne corporation manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96

    Structured Review

    Bio-Techne corporation hace2
    ( a ) Experimental setup. Antibody-sufficient (Ab + , n = 3-10) and antibody-deficient (Ab − , n = 3–28) <t>K18-hACE2</t> transgenic mice were infected with a target dose of 2 × 10 5 TCID 50 of SARS-CoV-2 D614G through aerosol exposure. Brain was collected and analyzed 9 days postchallenge. ( b ) Survival curve of Ab + ( n = 10, blue line) and Ab − mice upon infection ( n = 28, red line). (c) Quantification of SARS-CoV-2 RNA in the brain of the indicated mice. RNA values are expressed as copy number per ng of total RNA and the limit of detection is indicated as a dotted line. n = 3. ( d, e ) Experimental setup as described in Fig. . ( d ) Representative confocal immunofluorescence staining of lung sections from Ab + (upper panels) and Ab − (lower panels) mice 4 days after re-challenge. Cell nuclei are depicted in blue, B220 + cells in green and TCR-β + cells in white. Scale bar, 30 μm. ( e ) Representative flow cytometry plots (left) and frequency (right) of RBD-specific B cells in the mLN of indicated mice 4 days post re-challenge (pre-gated on live + /CD4 − / CD8 − / B220 + /CD19 + cells). n as indicated in Fig. . Data are expressed as mean ± SEM. Data are representative of at least 2 independent experiments. *p value < 0.05, **p value < 0.01, ***p value < 0.001; log-rank (Mantel-Cox) test ( b ); Two-tailed unpaired t-test ( c ); Kruskal-Wallis test followed by uncorrected Dunn’s test, each comparison stands alone ( e ).
    Hace2, supplied by Bio-Techne corporation, 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/result/hace2/product/Bio-Techne corporation
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hace2 - by Bioz Stars, 2024-05
    96/100 stars

    Images

    1) Product Images from "Antibody-independent protection against heterologous SARS-CoV-2 challenge conferred by prior infection or vaccination"

    Article Title: Antibody-independent protection against heterologous SARS-CoV-2 challenge conferred by prior infection or vaccination

    Journal: Nature Immunology

    doi: 10.1038/s41590-024-01787-z

    ( a ) Experimental setup. Antibody-sufficient (Ab + , n = 3-10) and antibody-deficient (Ab − , n = 3–28) K18-hACE2 transgenic mice were infected with a target dose of 2 × 10 5 TCID 50 of SARS-CoV-2 D614G through aerosol exposure. Brain was collected and analyzed 9 days postchallenge. ( b ) Survival curve of Ab + ( n = 10, blue line) and Ab − mice upon infection ( n = 28, red line). (c) Quantification of SARS-CoV-2 RNA in the brain of the indicated mice. RNA values are expressed as copy number per ng of total RNA and the limit of detection is indicated as a dotted line. n = 3. ( d, e ) Experimental setup as described in Fig. . ( d ) Representative confocal immunofluorescence staining of lung sections from Ab + (upper panels) and Ab − (lower panels) mice 4 days after re-challenge. Cell nuclei are depicted in blue, B220 + cells in green and TCR-β + cells in white. Scale bar, 30 μm. ( e ) Representative flow cytometry plots (left) and frequency (right) of RBD-specific B cells in the mLN of indicated mice 4 days post re-challenge (pre-gated on live + /CD4 − / CD8 − / B220 + /CD19 + cells). n as indicated in Fig. . Data are expressed as mean ± SEM. Data are representative of at least 2 independent experiments. *p value < 0.05, **p value < 0.01, ***p value < 0.001; log-rank (Mantel-Cox) test ( b ); Two-tailed unpaired t-test ( c ); Kruskal-Wallis test followed by uncorrected Dunn’s test, each comparison stands alone ( e ).
    Figure Legend Snippet: ( a ) Experimental setup. Antibody-sufficient (Ab + , n = 3-10) and antibody-deficient (Ab − , n = 3–28) K18-hACE2 transgenic mice were infected with a target dose of 2 × 10 5 TCID 50 of SARS-CoV-2 D614G through aerosol exposure. Brain was collected and analyzed 9 days postchallenge. ( b ) Survival curve of Ab + ( n = 10, blue line) and Ab − mice upon infection ( n = 28, red line). (c) Quantification of SARS-CoV-2 RNA in the brain of the indicated mice. RNA values are expressed as copy number per ng of total RNA and the limit of detection is indicated as a dotted line. n = 3. ( d, e ) Experimental setup as described in Fig. . ( d ) Representative confocal immunofluorescence staining of lung sections from Ab + (upper panels) and Ab − (lower panels) mice 4 days after re-challenge. Cell nuclei are depicted in blue, B220 + cells in green and TCR-β + cells in white. Scale bar, 30 μm. ( e ) Representative flow cytometry plots (left) and frequency (right) of RBD-specific B cells in the mLN of indicated mice 4 days post re-challenge (pre-gated on live + /CD4 − / CD8 − / B220 + /CD19 + cells). n as indicated in Fig. . Data are expressed as mean ± SEM. Data are representative of at least 2 independent experiments. *p value < 0.05, **p value < 0.01, ***p value < 0.001; log-rank (Mantel-Cox) test ( b ); Two-tailed unpaired t-test ( c ); Kruskal-Wallis test followed by uncorrected Dunn’s test, each comparison stands alone ( e ).

    Techniques Used: Transgenic Assay, Infection, Aerosol, Immunofluorescence, Staining, Flow Cytometry, Two Tailed Test, Comparison

    a , Experimental setup. Ab + ( n = 5) and Ab − ( n = 3–7) K18-hACE2 mice were primed with 2 × 10 5 TCID 50 of SARS-CoV-2 D614G and rechallenged with a higher dose (1 × 10 6 TCID 50 ) of SARS-CoV-2 B.1.1.529 (Omicron). Ab + ( n = 4–9) and Ab − ( n = 4–7) naïve mice, unexposed to the primary challenge, were infected with 1 × 10 6 TCID 50 of SARS-CoV-2 B.1.1.529. PBS-exposed mice were used as controls. Blood was collected 7, 14 and 21 days after the first infection. Blood, lung, NT and mediastinal lymph node (mLN) were collected 4 days after rechallenge. b , Anti-S1 RBD IgG levels in the plasma after the first challenge. c , d , SARS-CoV-2 RNA in the NT ( c ) and lung ( d ). RNA values as copy number per ng of total RNA and the LOD as a dashed line. e , Viral titers in the lung were determined by TCID 50 . f , Immunohistochemical micrographs of lung sections from PBS-, naïve- and primed-Ab + and Ab − mice. N-SARS-CoV-2-positive cells in brown. Scale bars, 100 μm. g – l , Flow cytometry plots ( g and j ), frequency ( h and k ) and absolute number ( i and l ) of CD8 + T cells ( g – i ) or CD4 + T cells ( j – l ) expressing IFN-γ and TNF in the lungs upon in vitro stimulation with a pool of SARS-CoV-2 peptides. Plots pregated as live + /B220 − /CD19 − /CD4 − /CD8 + ( g – i ) or CD8 − /CD4 + ( j – l ). m , Anti-S1 RBD IgG levels in the plasma 4 days after rechallenge. n , o , Flow cytometry plots ( n ) and frequency ( o ) of RBD-specific B cells detected by RBD-tetramers in the lungs (pregated on live + /CD4 − /CD8 − /B220 + /CD19 + ). p , q , Flow cytometry histogram ( p ) and geometric mean fluorescence intensity (gMFI) ( q ) of surface markers expressed by RBD-specific B cells in the lung of Ab + primed mice. As control, B cells negative for RBD-tetramer staining (gray). gMFI as log 2 (fold change) over control B cells. Data are expressed as mean ± s.e.m. and are representative of at least two independent experiments. Data in b – d are pooled from two independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001; Kruskal–Wallis test followed by uncorrected Dunn’s test; each comparison stands alone ( c – e , m and o ). Two-way ANOVA, Fisher’s LSD test (each comparison stands alone; b , h , i , k and l ). LSD, least significant difference.
    Figure Legend Snippet: a , Experimental setup. Ab + ( n = 5) and Ab − ( n = 3–7) K18-hACE2 mice were primed with 2 × 10 5 TCID 50 of SARS-CoV-2 D614G and rechallenged with a higher dose (1 × 10 6 TCID 50 ) of SARS-CoV-2 B.1.1.529 (Omicron). Ab + ( n = 4–9) and Ab − ( n = 4–7) naïve mice, unexposed to the primary challenge, were infected with 1 × 10 6 TCID 50 of SARS-CoV-2 B.1.1.529. PBS-exposed mice were used as controls. Blood was collected 7, 14 and 21 days after the first infection. Blood, lung, NT and mediastinal lymph node (mLN) were collected 4 days after rechallenge. b , Anti-S1 RBD IgG levels in the plasma after the first challenge. c , d , SARS-CoV-2 RNA in the NT ( c ) and lung ( d ). RNA values as copy number per ng of total RNA and the LOD as a dashed line. e , Viral titers in the lung were determined by TCID 50 . f , Immunohistochemical micrographs of lung sections from PBS-, naïve- and primed-Ab + and Ab − mice. N-SARS-CoV-2-positive cells in brown. Scale bars, 100 μm. g – l , Flow cytometry plots ( g and j ), frequency ( h and k ) and absolute number ( i and l ) of CD8 + T cells ( g – i ) or CD4 + T cells ( j – l ) expressing IFN-γ and TNF in the lungs upon in vitro stimulation with a pool of SARS-CoV-2 peptides. Plots pregated as live + /B220 − /CD19 − /CD4 − /CD8 + ( g – i ) or CD8 − /CD4 + ( j – l ). m , Anti-S1 RBD IgG levels in the plasma 4 days after rechallenge. n , o , Flow cytometry plots ( n ) and frequency ( o ) of RBD-specific B cells detected by RBD-tetramers in the lungs (pregated on live + /CD4 − /CD8 − /B220 + /CD19 + ). p , q , Flow cytometry histogram ( p ) and geometric mean fluorescence intensity (gMFI) ( q ) of surface markers expressed by RBD-specific B cells in the lung of Ab + primed mice. As control, B cells negative for RBD-tetramer staining (gray). gMFI as log 2 (fold change) over control B cells. Data are expressed as mean ± s.e.m. and are representative of at least two independent experiments. Data in b – d are pooled from two independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001; Kruskal–Wallis test followed by uncorrected Dunn’s test; each comparison stands alone ( c – e , m and o ). Two-way ANOVA, Fisher’s LSD test (each comparison stands alone; b , h , i , k and l ). LSD, least significant difference.

    Techniques Used: Infection, Immunohistochemical staining, Flow Cytometry, Expressing, In Vitro, Fluorescence, Staining, Comparison

    a , Amino acid sequence of human (h)ACE2 and mouse (m)ACE2. In red are the eight residues involved in the interaction with the SARS-CoV-2 spike protein. b , Molecular modeling of the interaction between SARS-CoV-2 spike RBD (orange) and hACE2 or mACE2 (cyan). Crosses indicate the absence of interaction. Electrostatic and hydrophobic interactions in green and yellow dashed lines. c , Experimental setup. The 3T3 cells transduced with lentiviral vectors to express hACE2 (blue symbols), mACE2 (gray symbols) and a hybrid human/mouse (hy)ACE2 (green symbols) were infected with different concentrations of SARS-CoV-2. Nontransduced (WT) 3T3 cells as control. n = 3 biological replicates. d , Dose-dependent viral activity in 3T3 cells infected with SARS-CoV-2 D614G (left), B.1.617.2 (middle) or B.1.1.529 (right). Infection rates as a percentage of the virus-induced cytopathic effect 72 h after infection. Comparison with WT 3T3 cells. n = 3 biological replicates. e , Design of human/mouse hybrid Ace2 allele. f , Experimental setup. K18-hACE2 transgenic mice ( n = 4) and hyACE2 knock-in mice ( n = 5) were infected with 5 × 10 5 TCID 50 of SARS-CoV-2 B.1.617.2 (Delta). PBS-exposed mice were used as controls ( n = 2). Peripheral blood, lung and NT were analyzed 6 days after challenge. g , SARS-CoV-2 RNA in the NT (left) and lung (right). RNA values as copy number per ng of total RNA and the LOD as a dashed line. h , Respiratory frequency (left) and Rpef (right) were assessed by WBP 5 days postinfection (average over a 15-min data collection period). i , Anti-S1 RBD IgG levels in the plasma. j , k , Absolute number of total CD8 + T cells ( j , left) and CD4 + T cells ( k , left) and of cytokine-producing CD8 + cells ( j , right) and CD4 + cells ( k , right) in the lung on in vitro stimulation with a pool of SARS-CoV-2 peptides. Data are expressed as mean ± s.e.m. and are representative of at least two independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001; Kruskal–Wallis test followed by uncorrected Dunn’s test; each comparison stands alone ( g – i , j and k (left)). Two-way ANOVA, Tukey’s multiple comparison ( j and k (right)); two-way ANOVA, Fisher’s LSD test (each comparison stands alone; d ).
    Figure Legend Snippet: a , Amino acid sequence of human (h)ACE2 and mouse (m)ACE2. In red are the eight residues involved in the interaction with the SARS-CoV-2 spike protein. b , Molecular modeling of the interaction between SARS-CoV-2 spike RBD (orange) and hACE2 or mACE2 (cyan). Crosses indicate the absence of interaction. Electrostatic and hydrophobic interactions in green and yellow dashed lines. c , Experimental setup. The 3T3 cells transduced with lentiviral vectors to express hACE2 (blue symbols), mACE2 (gray symbols) and a hybrid human/mouse (hy)ACE2 (green symbols) were infected with different concentrations of SARS-CoV-2. Nontransduced (WT) 3T3 cells as control. n = 3 biological replicates. d , Dose-dependent viral activity in 3T3 cells infected with SARS-CoV-2 D614G (left), B.1.617.2 (middle) or B.1.1.529 (right). Infection rates as a percentage of the virus-induced cytopathic effect 72 h after infection. Comparison with WT 3T3 cells. n = 3 biological replicates. e , Design of human/mouse hybrid Ace2 allele. f , Experimental setup. K18-hACE2 transgenic mice ( n = 4) and hyACE2 knock-in mice ( n = 5) were infected with 5 × 10 5 TCID 50 of SARS-CoV-2 B.1.617.2 (Delta). PBS-exposed mice were used as controls ( n = 2). Peripheral blood, lung and NT were analyzed 6 days after challenge. g , SARS-CoV-2 RNA in the NT (left) and lung (right). RNA values as copy number per ng of total RNA and the LOD as a dashed line. h , Respiratory frequency (left) and Rpef (right) were assessed by WBP 5 days postinfection (average over a 15-min data collection period). i , Anti-S1 RBD IgG levels in the plasma. j , k , Absolute number of total CD8 + T cells ( j , left) and CD4 + T cells ( k , left) and of cytokine-producing CD8 + cells ( j , right) and CD4 + cells ( k , right) in the lung on in vitro stimulation with a pool of SARS-CoV-2 peptides. Data are expressed as mean ± s.e.m. and are representative of at least two independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001; Kruskal–Wallis test followed by uncorrected Dunn’s test; each comparison stands alone ( g – i , j and k (left)). Two-way ANOVA, Tukey’s multiple comparison ( j and k (right)); two-way ANOVA, Fisher’s LSD test (each comparison stands alone; d ).

    Techniques Used: Sequencing, Transduction, Infection, Activity Assay, Virus, Comparison, Transgenic Assay, Knock-In, In Vitro

    ( a ) Amino acid sequence alignment of SARS-CoV-2 D614G, B.1.617.2 and B.1.1.529. The residues involved in the interaction with the human ACE2 are indicated in red. ( b ) Representative flow cytometry histograms representing the ACE2 expression by 3T3 cells and 3T3 cells transduced with murine ACE2, human ACE2 and hybrid ACE2. ( c, d ) SARS-CoV-2 titers in transduced 3T3 cells upon infection with D614G (left), B.1.617.2 (middle) or B.1.1.529 (right). In ( c ) titers were determined as percent of the virus-induced cytopathic effect evaluated 48 hours after infection. In ( d ) titers were determined by qPCR quantification of SARS-CoV-2 RNA in the supernatant 48 and 72 hours postinfection. n = 3 biological replicates. ( e ) Human ACE2 expression in the indicated organs from WT (gray), K18-hACE2 (blue) and hyACE2 (green) mice. Values were normalized to the reference gene Gapdh and expressed as fold increase over the limit of detection (LOD). n = 7-8 mice. ( f ) K18-hACE2 transgenic mice ( n = 5) and hyACE2 knock-in mice ( n = 5) were infected with a target dose of 2 ×10 5 or 5 ×10 5 TCID 50 of SARS-CoV-2 B.1.617.2 through aerosol exposure. Lung and nasal turbinates (NT) were collected and analyzed 3 days postchallenge. ( g , h ) Quantification of SARS-CoV-2 RNA in the NT ( g ) and in the lung ( h ) of the indicated mice. n as indicated in (f). RNA values are expressed as copy number per ng of total RNA and the limit of detection is indicated as a dotted line. Data are expressed as mean ± SEM. Data are representative of at least 2 independent experiments. *p value < 0.05, **p value < 0.01, ***p value < 0.001; Two-way ANOVA, Fisher’s LSD test (Each comparison stands alone) ( c – e , g , h ).
    Figure Legend Snippet: ( a ) Amino acid sequence alignment of SARS-CoV-2 D614G, B.1.617.2 and B.1.1.529. The residues involved in the interaction with the human ACE2 are indicated in red. ( b ) Representative flow cytometry histograms representing the ACE2 expression by 3T3 cells and 3T3 cells transduced with murine ACE2, human ACE2 and hybrid ACE2. ( c, d ) SARS-CoV-2 titers in transduced 3T3 cells upon infection with D614G (left), B.1.617.2 (middle) or B.1.1.529 (right). In ( c ) titers were determined as percent of the virus-induced cytopathic effect evaluated 48 hours after infection. In ( d ) titers were determined by qPCR quantification of SARS-CoV-2 RNA in the supernatant 48 and 72 hours postinfection. n = 3 biological replicates. ( e ) Human ACE2 expression in the indicated organs from WT (gray), K18-hACE2 (blue) and hyACE2 (green) mice. Values were normalized to the reference gene Gapdh and expressed as fold increase over the limit of detection (LOD). n = 7-8 mice. ( f ) K18-hACE2 transgenic mice ( n = 5) and hyACE2 knock-in mice ( n = 5) were infected with a target dose of 2 ×10 5 or 5 ×10 5 TCID 50 of SARS-CoV-2 B.1.617.2 through aerosol exposure. Lung and nasal turbinates (NT) were collected and analyzed 3 days postchallenge. ( g , h ) Quantification of SARS-CoV-2 RNA in the NT ( g ) and in the lung ( h ) of the indicated mice. n as indicated in (f). RNA values are expressed as copy number per ng of total RNA and the limit of detection is indicated as a dotted line. Data are expressed as mean ± SEM. Data are representative of at least 2 independent experiments. *p value < 0.05, **p value < 0.01, ***p value < 0.001; Two-way ANOVA, Fisher’s LSD test (Each comparison stands alone) ( c – e , g , h ).

    Techniques Used: Sequencing, Flow Cytometry, Expressing, Transduction, Infection, Virus, Transgenic Assay, Knock-In, Aerosol, Comparison

    anti ace2  (Bio-Techne corporation)


    Bioz Verified Symbol Bio-Techne corporation is a verified supplier
    Bioz Manufacturer Symbol Bio-Techne corporation manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96

    Structured Review

    Bio-Techne corporation anti ace2
    ( A ) (Top) Mean (+/− SEM) of SFV viral titers in NoDice FHA:DICER WT and N1 cells infected at an MOI of 1.10 −4 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Unpaired t-test with Welch’s correction. * p < 0.05. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in SFV infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( B ) (Top) Mean (+/− SEM) of EV71 viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 0.1 for 24 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test. ** p < 0.01. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in EV71 infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( C ) (Top) Mean (+/− SEM) of VSV-GFP viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 1.10 −5 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Unpaired t-test. ns: non-significant. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in VSV-GFP infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( D ) Western blot analysis of human <t>ACE2</t> and HA (for DICER) expression in NoDice FHA:DICER WT and N1 cells ( n = 3 biological replicates). Alpha-Tubulin was used as loading control. ( E ) Western blot analysis of DICER and NUCLEOCAPSID expression in SARS-CoV-2 infected NoDice FHA:DICER WT and N1 cells at an MOI of 0.001 for 48 h. GAPDH was used as loading control. Band intensity was quantified and normalized to GAPDH for three independent biological replicates, then represented as mean (+/− SD) under the corresponding lane. ( F ) Mean (+/− SEM) of SARS-CoV-2 viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 0.001 for 48 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test. * p < 0.05. .
    Anti Ace2, supplied by Bio-Techne corporation, 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/result/anti ace2/product/Bio-Techne corporation
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti ace2 - by Bioz Stars, 2024-05
    96/100 stars

    Images

    1) Product Images from "The helicase domain of human Dicer prevents RNAi-independent activation of antiviral and inflammatory pathways"

    Article Title: The helicase domain of human Dicer prevents RNAi-independent activation of antiviral and inflammatory pathways

    Journal: The EMBO Journal

    doi: 10.1038/s44318-024-00035-2

    ( A ) (Top) Mean (+/− SEM) of SFV viral titers in NoDice FHA:DICER WT and N1 cells infected at an MOI of 1.10 −4 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Unpaired t-test with Welch’s correction. * p < 0.05. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in SFV infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( B ) (Top) Mean (+/− SEM) of EV71 viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 0.1 for 24 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test. ** p < 0.01. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in EV71 infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( C ) (Top) Mean (+/− SEM) of VSV-GFP viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 1.10 −5 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Unpaired t-test. ns: non-significant. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in VSV-GFP infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( D ) Western blot analysis of human ACE2 and HA (for DICER) expression in NoDice FHA:DICER WT and N1 cells ( n = 3 biological replicates). Alpha-Tubulin was used as loading control. ( E ) Western blot analysis of DICER and NUCLEOCAPSID expression in SARS-CoV-2 infected NoDice FHA:DICER WT and N1 cells at an MOI of 0.001 for 48 h. GAPDH was used as loading control. Band intensity was quantified and normalized to GAPDH for three independent biological replicates, then represented as mean (+/− SD) under the corresponding lane. ( F ) Mean (+/− SEM) of SARS-CoV-2 viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 0.001 for 48 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test. * p < 0.05. .
    Figure Legend Snippet: ( A ) (Top) Mean (+/− SEM) of SFV viral titers in NoDice FHA:DICER WT and N1 cells infected at an MOI of 1.10 −4 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Unpaired t-test with Welch’s correction. * p < 0.05. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in SFV infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( B ) (Top) Mean (+/− SEM) of EV71 viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 0.1 for 24 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test. ** p < 0.01. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in EV71 infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( C ) (Top) Mean (+/− SEM) of VSV-GFP viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 1.10 −5 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Unpaired t-test. ns: non-significant. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in VSV-GFP infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( D ) Western blot analysis of human ACE2 and HA (for DICER) expression in NoDice FHA:DICER WT and N1 cells ( n = 3 biological replicates). Alpha-Tubulin was used as loading control. ( E ) Western blot analysis of DICER and NUCLEOCAPSID expression in SARS-CoV-2 infected NoDice FHA:DICER WT and N1 cells at an MOI of 0.001 for 48 h. GAPDH was used as loading control. Band intensity was quantified and normalized to GAPDH for three independent biological replicates, then represented as mean (+/− SD) under the corresponding lane. ( F ) Mean (+/− SEM) of SARS-CoV-2 viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 0.001 for 48 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test. * p < 0.05. .

    Techniques Used: Infection, Plaque Assay, Immunofluorescence, Staining, Western Blot, Expressing

    ( A ) Schematic representation of PKR and its two point mutants: K296R and T451A. ( B ) Western blot analysis of DICER, p-PKR, PKR, and CAPSID expression in SINV-GFP infected NoDice∆PKR FHA:DICER WT cells expressing MYC:EMPTY CTRL vector, MYC:PKR, MYC K296R or MYC:T451A at an MOI of 0.02 for 24 h. Alpha-Tubulin was used as loading control. Band intensity was quantified and normalized to Tubulin for three independent biological replicates, then represented as mean (+/− SD) under the corresponding lane; p-PKR = p-PKR/PKR quantification. ( C ) Mean (+/− SEM) of SINV-GFP viral titers in the same samples as in (B), infected at an MOI of 0.02 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Ordinary one-way ANOVA test with Dunnett’s correction. ns: non-significant. ( D ) Western blot analysis of DICER, p-PKR, PKR and CAPSID expression in SINV-GFP infected NoDice∆PKR FHA:DICER N1 cells expressing MYC:EMPTY CTRL vector, MYC:PKR, MYC K296R or MYC:T451A at an MOI of 0.02 for 24 h. Alpha-Tubulin was used as loading control. Band intensity was quantified and normalized to Tubulin for three independent biological replicates, then represented as mean (+/− SD) under the corresponding lane; p-PKR = p-PKR/PKR quantification. ( E ) Mean (+/− SEM) of SINV-GFP viral titers in the same samples as in ( D ), infected at an MOI of 0.02 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Ordinary one-way ANOVA test with Dunnett’s correction. * p < 0.05. ( F ) Western blot analysis of DICER, human ACE2, PKR and NUCLEOCAPSID expression in SARS-CoV-2 infected NoDice FHA:DICER WT or N1 and NoDice∆PKR FHA:DICER WT or N1 cells at an MOI of 0.001 for 48 h. GAPDH was used as loading control. Band intensity was quantified and normalized to GAPDH for three independent biological replicates, then represented as mean (+/− SD) under the corresponding lane. ( G ) Mean (+/− SEM) of SARS-CoV-2 viral titers in the same samples as in ( F ), infected at an MOI of 0.001 for 48 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test with Welch’s correction. * p < 0.05; ** p < 0.01. .
    Figure Legend Snippet: ( A ) Schematic representation of PKR and its two point mutants: K296R and T451A. ( B ) Western blot analysis of DICER, p-PKR, PKR, and CAPSID expression in SINV-GFP infected NoDice∆PKR FHA:DICER WT cells expressing MYC:EMPTY CTRL vector, MYC:PKR, MYC K296R or MYC:T451A at an MOI of 0.02 for 24 h. Alpha-Tubulin was used as loading control. Band intensity was quantified and normalized to Tubulin for three independent biological replicates, then represented as mean (+/− SD) under the corresponding lane; p-PKR = p-PKR/PKR quantification. ( C ) Mean (+/− SEM) of SINV-GFP viral titers in the same samples as in (B), infected at an MOI of 0.02 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Ordinary one-way ANOVA test with Dunnett’s correction. ns: non-significant. ( D ) Western blot analysis of DICER, p-PKR, PKR and CAPSID expression in SINV-GFP infected NoDice∆PKR FHA:DICER N1 cells expressing MYC:EMPTY CTRL vector, MYC:PKR, MYC K296R or MYC:T451A at an MOI of 0.02 for 24 h. Alpha-Tubulin was used as loading control. Band intensity was quantified and normalized to Tubulin for three independent biological replicates, then represented as mean (+/− SD) under the corresponding lane; p-PKR = p-PKR/PKR quantification. ( E ) Mean (+/− SEM) of SINV-GFP viral titers in the same samples as in ( D ), infected at an MOI of 0.02 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Ordinary one-way ANOVA test with Dunnett’s correction. * p < 0.05. ( F ) Western blot analysis of DICER, human ACE2, PKR and NUCLEOCAPSID expression in SARS-CoV-2 infected NoDice FHA:DICER WT or N1 and NoDice∆PKR FHA:DICER WT or N1 cells at an MOI of 0.001 for 48 h. GAPDH was used as loading control. Band intensity was quantified and normalized to GAPDH for three independent biological replicates, then represented as mean (+/− SD) under the corresponding lane. ( G ) Mean (+/− SEM) of SARS-CoV-2 viral titers in the same samples as in ( F ), infected at an MOI of 0.001 for 48 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test with Welch’s correction. * p < 0.05; ** p < 0.01. .

    Techniques Used: Western Blot, Expressing, Infection, Plasmid Preparation, Plaque Assay

    ace2 antibody  (Bio-Techne corporation)


    Bioz Verified Symbol Bio-Techne corporation is a verified supplier
    Bioz Manufacturer Symbol Bio-Techne corporation manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Bio-Techne corporation ace2 antibody
    Ace2 Antibody, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ace2 antibody/product/Bio-Techne corporation
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ace2 antibody - by Bioz Stars, 2024-05
    94/100 stars

    Images

    ace2 antibody  (Bio-Techne corporation)


    Bioz Verified Symbol Bio-Techne corporation is a verified supplier
    Bioz Manufacturer Symbol Bio-Techne corporation manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Bio-Techne corporation ace2 antibody
    Primer sequences for RT-PCR.
    Ace2 Antibody, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ace2 antibody/product/Bio-Techne corporation
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ace2 antibody - by Bioz Stars, 2024-05
    94/100 stars

    Images

    1) Product Images from "Extracellular Vesicles and Their Renin–Angiotensin Cargo as a Link between Metabolic Syndrome and Parkinson’s Disease"

    Article Title: Extracellular Vesicles and Their Renin–Angiotensin Cargo as a Link between Metabolic Syndrome and Parkinson’s Disease

    Journal: Antioxidants

    doi: 10.3390/antiox12122045

    Primer sequences for RT-PCR.
    Figure Legend Snippet: Primer sequences for RT-PCR.

    Techniques Used:

    mRNA-cargo of RAS components in EVs. Expression of pro-inflammatory components: AGT ( A ), AT1 ( B ), and PRR ( C ) was increased in EV MetS relative to EV Control , which was inhibited when MetS animals were treated with the AT1 receptor blocker candesartan. The expression of the anti-inflammatory component MasR ( E ) was increased in EV MetS+CAND . For mRNA, the comparative cycle threshold values method (2− ΔΔCt ) was used. Data are given as the mean ± SEM. * p < 0.05 compared to the EV Control group; # p < 0.05 compared to the EV MetS group (Kruskal–Wallis one-way ANOVA with the Student-Newman-Keuls Method as a post hoc test ( A ); one-way ANOVA with the Student-Newman-Keuls Method as a post hoc test ( B – E )). ACE2: angiotensin-converting enzyme 2; AGT: angiotensinogen; AT1: angiotensin type 1 receptor, CAND: candesartan; EVs: extracellular vesicles; EV Control : EVs isolated from the serum of control animals; EV MetS : EVs isolated from the serum of metabolic syndrome animals; EV MetS+CAND : EVs isolated from the serum of metabolic syndrome animals treated with candesartan; MasR: Mas-related receptor; MetS: metabolic syndrome; PRR: (pro)-renin receptor.
    Figure Legend Snippet: mRNA-cargo of RAS components in EVs. Expression of pro-inflammatory components: AGT ( A ), AT1 ( B ), and PRR ( C ) was increased in EV MetS relative to EV Control , which was inhibited when MetS animals were treated with the AT1 receptor blocker candesartan. The expression of the anti-inflammatory component MasR ( E ) was increased in EV MetS+CAND . For mRNA, the comparative cycle threshold values method (2− ΔΔCt ) was used. Data are given as the mean ± SEM. * p < 0.05 compared to the EV Control group; # p < 0.05 compared to the EV MetS group (Kruskal–Wallis one-way ANOVA with the Student-Newman-Keuls Method as a post hoc test ( A ); one-way ANOVA with the Student-Newman-Keuls Method as a post hoc test ( B – E )). ACE2: angiotensin-converting enzyme 2; AGT: angiotensinogen; AT1: angiotensin type 1 receptor, CAND: candesartan; EVs: extracellular vesicles; EV Control : EVs isolated from the serum of control animals; EV MetS : EVs isolated from the serum of metabolic syndrome animals; EV MetS+CAND : EVs isolated from the serum of metabolic syndrome animals treated with candesartan; MasR: Mas-related receptor; MetS: metabolic syndrome; PRR: (pro)-renin receptor.

    Techniques Used: Expressing, Isolation

    Analysis of EVs from rat serum using SP-IRIS/ExoView ® . An increase in the fluorescence intensity of the pro-inflammatory AT1 receptor ( A ) and a non-significant trend towards increasing the fluorescence intensity of PRR ( B ) was observed in EV MetS relative to EV Control captured by tetraspanins (i.e., total serum EVs). Similarly, a significant increase in AT1 ( C ) and a non-significant increase in PRR ( D ) fluorescence was observed in EV MetS when captured by Cav + Perlip (i.e., EVs from fat tissue). A tendency to decrease ACE2 levels ( E , G ), and a significant decrease in the levels of the anti-inflammatory Mas receptor was observed in EV MetS relative to EV Control both captured by tetraspanins ( E , F ) and by Cav + Perlip ( G , H ). Interestingly, EVs from MetS rats treated with candesartan (EV MetS+CAND ) showed a significant increase in the fluorescence intensity of the anti-inflammatory components ACE2 ( E , G ) and MasR ( F , H ). Photographs ( I – K ) show label-free interferometry (IFM) images of a representative anti-Caveolin capture chip from control rats ( I ), MetS rats ( J ), and MetS rats treated with candesartan ( K ). Pink (AT1 receptor), green (PRR), blue (ACE2), and red (MasR) fluorescence can be observed. Scale bars: 10 µm. Data are given as the mean ± SEM. * p < 0.05 compared to the EV Control group; # p < 0.05 compared to the EV MetS group (Kruskal–Wallis one-way ANOVA with the Student-Newman-Keuls method as a post hoc test ( A , E , F ); one-way ANOVA with the Student-Newman-Keuls method as a post hoc test ( B – D , G , H )). ACE2: angiotensin-converting enzyme 2; AT1: angiotensin type 1 receptor; CAND: candesartan; Cav: Caveolin; EVs: extracellular vesicles; EV Control : EVs isolated from the serum of control animals; EV MetS : EVs isolated from the serum of metabolic syndrome animals; EV MetS+CAND : EVs isolated from the serum of metabolic syndrome animals treated with candesartan; MasR: Mas-related receptor; MetS: metabolic syndrome; Perilip: Perilipin-1, PRR: (pro)-renin receptor.
    Figure Legend Snippet: Analysis of EVs from rat serum using SP-IRIS/ExoView ® . An increase in the fluorescence intensity of the pro-inflammatory AT1 receptor ( A ) and a non-significant trend towards increasing the fluorescence intensity of PRR ( B ) was observed in EV MetS relative to EV Control captured by tetraspanins (i.e., total serum EVs). Similarly, a significant increase in AT1 ( C ) and a non-significant increase in PRR ( D ) fluorescence was observed in EV MetS when captured by Cav + Perlip (i.e., EVs from fat tissue). A tendency to decrease ACE2 levels ( E , G ), and a significant decrease in the levels of the anti-inflammatory Mas receptor was observed in EV MetS relative to EV Control both captured by tetraspanins ( E , F ) and by Cav + Perlip ( G , H ). Interestingly, EVs from MetS rats treated with candesartan (EV MetS+CAND ) showed a significant increase in the fluorescence intensity of the anti-inflammatory components ACE2 ( E , G ) and MasR ( F , H ). Photographs ( I – K ) show label-free interferometry (IFM) images of a representative anti-Caveolin capture chip from control rats ( I ), MetS rats ( J ), and MetS rats treated with candesartan ( K ). Pink (AT1 receptor), green (PRR), blue (ACE2), and red (MasR) fluorescence can be observed. Scale bars: 10 µm. Data are given as the mean ± SEM. * p < 0.05 compared to the EV Control group; # p < 0.05 compared to the EV MetS group (Kruskal–Wallis one-way ANOVA with the Student-Newman-Keuls method as a post hoc test ( A , E , F ); one-way ANOVA with the Student-Newman-Keuls method as a post hoc test ( B – D , G , H )). ACE2: angiotensin-converting enzyme 2; AT1: angiotensin type 1 receptor; CAND: candesartan; Cav: Caveolin; EVs: extracellular vesicles; EV Control : EVs isolated from the serum of control animals; EV MetS : EVs isolated from the serum of metabolic syndrome animals; EV MetS+CAND : EVs isolated from the serum of metabolic syndrome animals treated with candesartan; MasR: Mas-related receptor; MetS: metabolic syndrome; Perilip: Perilipin-1, PRR: (pro)-renin receptor.

    Techniques Used: Fluorescence, Isolation

    RAS dysregulation induced by EV MetS in the N27 dopaminergic cell line and the C6 astrocytic cell line. The increase in the mRNA expression of the pro-inflammatory RAS components AGT ( A , F ), AT1 ( B , G ), and PRR ( C , H ) induced by EV MetS uptaking was inhibited when cells were treated with candesartan. The mRNA expression of the anti-inflammatory RAS components ACE2 ( D , I ) and MasR ( E , J ) was increased after candesartan treatment. For mRNA, the comparative cycle threshold values method (2− ΔΔCt ) was used. Data are given as the mean ± SEM. * p < 0.05 compared to the control group; # p < 0.05 compared to the EV MetS group (one-way ANOVA with the Student-Newman-Keuls Method as a post hoc test ( A – D , F , H – J ); Kruskal–Wallis one-way ANOVA with the Student-Newman-Keuls method ( E ) and Dunn’s method ( G ) as post hoc tests). ACE2: angiotensin-converting enzyme 2; AGT: angiotensinogen; AT1: angiotensin type 1 receptor; CAND: candesartan; EVs: extracellular vesicles; EV MetS : EVs isolated from the serum of metabolic syndrome animals; MasR: Mas-related receptor; MetS: metabolic syndrome; PRR: (pro)-renin receptor.
    Figure Legend Snippet: RAS dysregulation induced by EV MetS in the N27 dopaminergic cell line and the C6 astrocytic cell line. The increase in the mRNA expression of the pro-inflammatory RAS components AGT ( A , F ), AT1 ( B , G ), and PRR ( C , H ) induced by EV MetS uptaking was inhibited when cells were treated with candesartan. The mRNA expression of the anti-inflammatory RAS components ACE2 ( D , I ) and MasR ( E , J ) was increased after candesartan treatment. For mRNA, the comparative cycle threshold values method (2− ΔΔCt ) was used. Data are given as the mean ± SEM. * p < 0.05 compared to the control group; # p < 0.05 compared to the EV MetS group (one-way ANOVA with the Student-Newman-Keuls Method as a post hoc test ( A – D , F , H – J ); Kruskal–Wallis one-way ANOVA with the Student-Newman-Keuls method ( E ) and Dunn’s method ( G ) as post hoc tests). ACE2: angiotensin-converting enzyme 2; AGT: angiotensinogen; AT1: angiotensin type 1 receptor; CAND: candesartan; EVs: extracellular vesicles; EV MetS : EVs isolated from the serum of metabolic syndrome animals; MasR: Mas-related receptor; MetS: metabolic syndrome; PRR: (pro)-renin receptor.

    Techniques Used: Expressing, Isolation

    antibodies to ace2  (Bio-Techne corporation)


    Bioz Verified Symbol Bio-Techne corporation is a verified supplier
    Bioz Manufacturer Symbol Bio-Techne corporation manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 93

    Structured Review

    Bio-Techne corporation antibodies to ace2
    Antibodies To Ace2, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/antibodies to ace2/product/Bio-Techne corporation
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    antibodies to ace2 - by Bioz Stars, 2024-05
    93/100 stars

    Images

    anti ace2  (Bio-Techne corporation)


    Bioz Verified Symbol Bio-Techne corporation is a verified supplier
    Bioz Manufacturer Symbol Bio-Techne corporation manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96

    Structured Review

    Bio-Techne corporation anti ace2
    A, (Top) Mean (+/-SEM) of SFV viral titers in NoDice FHA:DICER WT #4 and N1 #6 infected at an MOI of 1.10 -4 for 24h (n = 3) from plaque assay quantification. Unpaired t-test. *: p < 0.05. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT #4 and N1 #6 cells in SFV infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Magnification 40X; scale bar = 100 µm. B, (Top) Mean (+/-SEM) of EV71 viral titers in NoDice FHA:DICER WT #4 and N1 #6 infected at an MOI of 0.1 for 24h (n = 3) from TCID50 quantification. Unpaired t-test. *: p < 0.05. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT #4 and N1 #6 cells in EV71 infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Magnification 40X; scale bar = 100 µm. C, (Top) Mean (+/-SEM) of VSV-GFP viral titers in NoDice FHA:DICER WT #4 and N1 #6 infected at an MOI of 1.10 -5 for 24h (n = 3) from plaque assay quantification. Unpaired t-test. ns: non-significant. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT #4 and N1 #6 cells in VSV-GFP infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Magnification 40X; scale bar = 100 µm. D, Western blot analysis of human <t>ACE2</t> expression in NoDice FHA:DICER WT #4 and N1 #6. Gamma-Tubulin was used as a loading control. E, Western blot analysis of DICER, p-PKR, PKR and NUCLEOCAPSID expression in SARS-CoV-2 infected NoDice FHA:DICER WT #4 and N1 #6 cells at an MOI of 0.001 for 48h. GAPDH was used as loading control. F, Mean (+/-SEM) of SARS-CoV-2 viral titers in NoDice FHA:DICER WT #4 and N1 #6 infected at an MOI of 0.001 for 48h (n = 3) from TCID50 quantification. Unpaired t-test. *: p < 0.05. G, Immunofluorescence analysis on NoDice FHA:DICER WT #4 and N1 #6 cells in SARS-CoV-2 infected cells at an MOI of 0.001 for 48h. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Magnification 40X; scale bar = 100 µm. H, RT-qPCR on SARS-CoV-2 genome (ORF1A and SPIKE) in NoDice FHA:DICER WT #4 and N1 #6 cells infected with SARS-CoV-2 at an MOI of 0.001 for 48h. Mean (+/-SEM); n = 3. Unpaired t-test. *: p < 0.05; ns: non-significant.
    Anti Ace2, supplied by Bio-Techne corporation, 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/result/anti ace2/product/Bio-Techne corporation
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti ace2 - by Bioz Stars, 2024-05
    96/100 stars

    Images

    1) Product Images from "Canonical and non-canonical contributions of human Dicer helicase domain in antiviral defense"

    Article Title: Canonical and non-canonical contributions of human Dicer helicase domain in antiviral defense

    Journal: bioRxiv

    doi: 10.1101/2023.08.23.554407

    A, (Top) Mean (+/-SEM) of SFV viral titers in NoDice FHA:DICER WT #4 and N1 #6 infected at an MOI of 1.10 -4 for 24h (n = 3) from plaque assay quantification. Unpaired t-test. *: p < 0.05. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT #4 and N1 #6 cells in SFV infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Magnification 40X; scale bar = 100 µm. B, (Top) Mean (+/-SEM) of EV71 viral titers in NoDice FHA:DICER WT #4 and N1 #6 infected at an MOI of 0.1 for 24h (n = 3) from TCID50 quantification. Unpaired t-test. *: p < 0.05. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT #4 and N1 #6 cells in EV71 infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Magnification 40X; scale bar = 100 µm. C, (Top) Mean (+/-SEM) of VSV-GFP viral titers in NoDice FHA:DICER WT #4 and N1 #6 infected at an MOI of 1.10 -5 for 24h (n = 3) from plaque assay quantification. Unpaired t-test. ns: non-significant. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT #4 and N1 #6 cells in VSV-GFP infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Magnification 40X; scale bar = 100 µm. D, Western blot analysis of human ACE2 expression in NoDice FHA:DICER WT #4 and N1 #6. Gamma-Tubulin was used as a loading control. E, Western blot analysis of DICER, p-PKR, PKR and NUCLEOCAPSID expression in SARS-CoV-2 infected NoDice FHA:DICER WT #4 and N1 #6 cells at an MOI of 0.001 for 48h. GAPDH was used as loading control. F, Mean (+/-SEM) of SARS-CoV-2 viral titers in NoDice FHA:DICER WT #4 and N1 #6 infected at an MOI of 0.001 for 48h (n = 3) from TCID50 quantification. Unpaired t-test. *: p < 0.05. G, Immunofluorescence analysis on NoDice FHA:DICER WT #4 and N1 #6 cells in SARS-CoV-2 infected cells at an MOI of 0.001 for 48h. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Magnification 40X; scale bar = 100 µm. H, RT-qPCR on SARS-CoV-2 genome (ORF1A and SPIKE) in NoDice FHA:DICER WT #4 and N1 #6 cells infected with SARS-CoV-2 at an MOI of 0.001 for 48h. Mean (+/-SEM); n = 3. Unpaired t-test. *: p < 0.05; ns: non-significant.
    Figure Legend Snippet: A, (Top) Mean (+/-SEM) of SFV viral titers in NoDice FHA:DICER WT #4 and N1 #6 infected at an MOI of 1.10 -4 for 24h (n = 3) from plaque assay quantification. Unpaired t-test. *: p < 0.05. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT #4 and N1 #6 cells in SFV infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Magnification 40X; scale bar = 100 µm. B, (Top) Mean (+/-SEM) of EV71 viral titers in NoDice FHA:DICER WT #4 and N1 #6 infected at an MOI of 0.1 for 24h (n = 3) from TCID50 quantification. Unpaired t-test. *: p < 0.05. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT #4 and N1 #6 cells in EV71 infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Magnification 40X; scale bar = 100 µm. C, (Top) Mean (+/-SEM) of VSV-GFP viral titers in NoDice FHA:DICER WT #4 and N1 #6 infected at an MOI of 1.10 -5 for 24h (n = 3) from plaque assay quantification. Unpaired t-test. ns: non-significant. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT #4 and N1 #6 cells in VSV-GFP infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Magnification 40X; scale bar = 100 µm. D, Western blot analysis of human ACE2 expression in NoDice FHA:DICER WT #4 and N1 #6. Gamma-Tubulin was used as a loading control. E, Western blot analysis of DICER, p-PKR, PKR and NUCLEOCAPSID expression in SARS-CoV-2 infected NoDice FHA:DICER WT #4 and N1 #6 cells at an MOI of 0.001 for 48h. GAPDH was used as loading control. F, Mean (+/-SEM) of SARS-CoV-2 viral titers in NoDice FHA:DICER WT #4 and N1 #6 infected at an MOI of 0.001 for 48h (n = 3) from TCID50 quantification. Unpaired t-test. *: p < 0.05. G, Immunofluorescence analysis on NoDice FHA:DICER WT #4 and N1 #6 cells in SARS-CoV-2 infected cells at an MOI of 0.001 for 48h. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Magnification 40X; scale bar = 100 µm. H, RT-qPCR on SARS-CoV-2 genome (ORF1A and SPIKE) in NoDice FHA:DICER WT #4 and N1 #6 cells infected with SARS-CoV-2 at an MOI of 0.001 for 48h. Mean (+/-SEM); n = 3. Unpaired t-test. *: p < 0.05; ns: non-significant.

    Techniques Used: Infection, Plaque Assay, Immunofluorescence, Staining, Western Blot, Expressing, Quantitative RT-PCR

    goat anti ace2  (Bio-Techne corporation)


    Bioz Verified Symbol Bio-Techne corporation is a verified supplier
    Bioz Manufacturer Symbol Bio-Techne corporation manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96

    Structured Review

    Bio-Techne corporation goat anti ace2
    Goat Anti Ace2, supplied by Bio-Techne corporation, 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/result/goat anti ace2/product/Bio-Techne corporation
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti ace2 - by Bioz Stars, 2024-05
    96/100 stars

    Images

    goat anti ace2  (Bio-Techne corporation)


    Bioz Verified Symbol Bio-Techne corporation is a verified supplier
    Bioz Manufacturer Symbol Bio-Techne corporation manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96

    Structured Review

    Bio-Techne corporation goat anti ace2
    The characteristics of the IEC#17. The IEC#17 monolayers were cultured in a differentiation medium from day 2. (A, B) IEC#17 cells were collected at the indicated time points. Relative mRNA expression of the indicated genes in IEC#17 during the course of differentiation (days 2, 4, and 6) were determined by qRT-PCR and normalized against the expression of GAPDH . Each result was normalized by the expression level at two-day after differentiation. Each value is representative of at least three independent experiments and is shown as the mean ± SD from three wells of cells of each culture group. The significant differences were determined using one-way ANOVA. (A) The transcription of the major intestinal markers was measured using qRT-PCR. We included LYZ , SI , CHGA , VIL1 , LGR5 and MUC2 which is the marker of Paneth cell, enterocyte, enteroendocrine cell, IEC, stem cell and goblet cell, respectively. (B) The transcription of the major host factors important for SARS-CoV-2 infection was measured using qRT-PCR. We included <t>ACE2</t> , FURIN , CTSL , TMPRSS2 and TMPRSS4 . (C) Schematic diagrams of the immunostaining analysis. The black arrow indicates the direction from which the photograph was taken. (D) The protein expressions of ZO-1, ACE2, and TMPRSS2 in IEC#17 monolayers were visualized by immunofluorescence. IEC#17 monolayers were fixed and stained at the indicated time points. The scale bar indicates 50 µm. (E) The protein expressions of ACE2 in IEC#17 monolayers with polarity were visualized by immunofluorescence. The photo was taken from vertical angle at six-day after differentiation. The white broken bar indicates Transwell membranes. *0.01 < P < 0.05; **0.005 < P < 0.01; *** P < 0.005.
    Goat Anti Ace2, supplied by Bio-Techne corporation, 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/result/goat anti ace2/product/Bio-Techne corporation
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti ace2 - by Bioz Stars, 2024-05
    96/100 stars

    Images

    1) Product Images from "Effective SARS-CoV-2 replication of monolayers of intestinal epithelial cells differentiated from human induced pluripotent stem cells"

    Article Title: Effective SARS-CoV-2 replication of monolayers of intestinal epithelial cells differentiated from human induced pluripotent stem cells

    Journal: Scientific Reports

    doi: 10.1038/s41598-023-38548-1

    The characteristics of the IEC#17. The IEC#17 monolayers were cultured in a differentiation medium from day 2. (A, B) IEC#17 cells were collected at the indicated time points. Relative mRNA expression of the indicated genes in IEC#17 during the course of differentiation (days 2, 4, and 6) were determined by qRT-PCR and normalized against the expression of GAPDH . Each result was normalized by the expression level at two-day after differentiation. Each value is representative of at least three independent experiments and is shown as the mean ± SD from three wells of cells of each culture group. The significant differences were determined using one-way ANOVA. (A) The transcription of the major intestinal markers was measured using qRT-PCR. We included LYZ , SI , CHGA , VIL1 , LGR5 and MUC2 which is the marker of Paneth cell, enterocyte, enteroendocrine cell, IEC, stem cell and goblet cell, respectively. (B) The transcription of the major host factors important for SARS-CoV-2 infection was measured using qRT-PCR. We included ACE2 , FURIN , CTSL , TMPRSS2 and TMPRSS4 . (C) Schematic diagrams of the immunostaining analysis. The black arrow indicates the direction from which the photograph was taken. (D) The protein expressions of ZO-1, ACE2, and TMPRSS2 in IEC#17 monolayers were visualized by immunofluorescence. IEC#17 monolayers were fixed and stained at the indicated time points. The scale bar indicates 50 µm. (E) The protein expressions of ACE2 in IEC#17 monolayers with polarity were visualized by immunofluorescence. The photo was taken from vertical angle at six-day after differentiation. The white broken bar indicates Transwell membranes. *0.01 < P < 0.05; **0.005 < P < 0.01; *** P < 0.005.
    Figure Legend Snippet: The characteristics of the IEC#17. The IEC#17 monolayers were cultured in a differentiation medium from day 2. (A, B) IEC#17 cells were collected at the indicated time points. Relative mRNA expression of the indicated genes in IEC#17 during the course of differentiation (days 2, 4, and 6) were determined by qRT-PCR and normalized against the expression of GAPDH . Each result was normalized by the expression level at two-day after differentiation. Each value is representative of at least three independent experiments and is shown as the mean ± SD from three wells of cells of each culture group. The significant differences were determined using one-way ANOVA. (A) The transcription of the major intestinal markers was measured using qRT-PCR. We included LYZ , SI , CHGA , VIL1 , LGR5 and MUC2 which is the marker of Paneth cell, enterocyte, enteroendocrine cell, IEC, stem cell and goblet cell, respectively. (B) The transcription of the major host factors important for SARS-CoV-2 infection was measured using qRT-PCR. We included ACE2 , FURIN , CTSL , TMPRSS2 and TMPRSS4 . (C) Schematic diagrams of the immunostaining analysis. The black arrow indicates the direction from which the photograph was taken. (D) The protein expressions of ZO-1, ACE2, and TMPRSS2 in IEC#17 monolayers were visualized by immunofluorescence. IEC#17 monolayers were fixed and stained at the indicated time points. The scale bar indicates 50 µm. (E) The protein expressions of ACE2 in IEC#17 monolayers with polarity were visualized by immunofluorescence. The photo was taken from vertical angle at six-day after differentiation. The white broken bar indicates Transwell membranes. *0.01 < P < 0.05; **0.005 < P < 0.01; *** P < 0.005.

    Techniques Used: Cell Culture, Expressing, Quantitative RT-PCR, Marker, Infection, Immunostaining, Immunofluorescence, Staining

    The potential of IEC#17 for the efficient SARS-CoV-2 infection. (A) IEC#17 and Vero cells were seeded in a plate and infected with SARS-CoV-2 at an MOI of 0.1. The supernatants were collected at indicated time points. SARS-CoV-2 growth was measured by determining the TCID 50 on VeroE6/TMPRSS2 cells. Each value is representative of at least three independent experiments and is shown as the mean ± SD from three wells of supernatants of each culture group. The significant differences were determined using two-way ANOVA. (B – D) The transmission electron microscopic analysis of IEC#17 monolayers infected with SARS-CoV-2. IEC#17 cells were seeded on Transwell membranes (C) or Cell Desks (D) . Each colored squares indicate each magnified panels. (B) Schematic diagrams of the transmission electron microscopic analysis. The black arrow indicates the direction from which the photograph was taken. (C) The IEC#17 were cultured to harbor the polarity and infected with SARS-CoV-2 at an MOI of 0.1 from apical side. At 24 hpi, infected cells were fixed and analyzed. The photos were taken from vertical angle. The black and white arrows indicate SARS-CoV-2 particle and viral replication organelles (ROs), respectively. The scale bars indicate 0.5 µm. (D) The IEC#17 were cultured by the normal method (no polarity) and infected with SARS-CoV-2 at an MOI of 0.1. At 24 hpi, infected cells were fixed and analyzed. The photos were taken from horizontal angles. The black arrows indicate SARS-CoV-2 particle. The scale bars indicate 0.5 µm. (E , F) Immunofluorescence analysis of (E) SARS-CoV-2 NP and SP, or (F) ACE2, TMPRSS2, and SARS-CoV-2 SP in IEC#17 monolayers infected with SARS-CoV-2. The photo was taken from horizontal angles at 24 hpi. The scale bars indicate 50 µm.
    Figure Legend Snippet: The potential of IEC#17 for the efficient SARS-CoV-2 infection. (A) IEC#17 and Vero cells were seeded in a plate and infected with SARS-CoV-2 at an MOI of 0.1. The supernatants were collected at indicated time points. SARS-CoV-2 growth was measured by determining the TCID 50 on VeroE6/TMPRSS2 cells. Each value is representative of at least three independent experiments and is shown as the mean ± SD from three wells of supernatants of each culture group. The significant differences were determined using two-way ANOVA. (B – D) The transmission electron microscopic analysis of IEC#17 monolayers infected with SARS-CoV-2. IEC#17 cells were seeded on Transwell membranes (C) or Cell Desks (D) . Each colored squares indicate each magnified panels. (B) Schematic diagrams of the transmission electron microscopic analysis. The black arrow indicates the direction from which the photograph was taken. (C) The IEC#17 were cultured to harbor the polarity and infected with SARS-CoV-2 at an MOI of 0.1 from apical side. At 24 hpi, infected cells were fixed and analyzed. The photos were taken from vertical angle. The black and white arrows indicate SARS-CoV-2 particle and viral replication organelles (ROs), respectively. The scale bars indicate 0.5 µm. (D) The IEC#17 were cultured by the normal method (no polarity) and infected with SARS-CoV-2 at an MOI of 0.1. At 24 hpi, infected cells were fixed and analyzed. The photos were taken from horizontal angles. The black arrows indicate SARS-CoV-2 particle. The scale bars indicate 0.5 µm. (E , F) Immunofluorescence analysis of (E) SARS-CoV-2 NP and SP, or (F) ACE2, TMPRSS2, and SARS-CoV-2 SP in IEC#17 monolayers infected with SARS-CoV-2. The photo was taken from horizontal angles at 24 hpi. The scale bars indicate 50 µm.

    Techniques Used: Infection, Transmission Assay, Cell Culture, Immunofluorescence

    ace2 goat  (Bio-Techne corporation)


    Bioz Verified Symbol Bio-Techne corporation is a verified supplier
    Bioz Manufacturer Symbol Bio-Techne corporation manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96

    Structured Review

    Bio-Techne corporation ace2 goat
    Characterization of iPSC-derived neurons and astrocytes. (A) Representative images of immunocytochemical staining of iPSC-derived NGN2-neurons with MAP2, TUBB3, GABA, CUX1, and VGLUT1 antibodies. Scale bar, 50 μm. (B) Representative images of immunocytochemical staining of iPSC-derived astrocytes with GFAP, S100β, and AQP4 antibodies. Scale bar, 100 μm. (C) Mean normalized mRNA expression of astrocyte markers GFAP and S100β in iPSCs and iPSC-derived astrocytes. Values are normalized to human GAPDH expression. N = 4 cell lines. (D) Quantification of the mean firing rate (in hertz), percentage of electrodes partaking in bursts, percentage of electrodes partaking in network bursts, and network burst duration (in seconds) in iPCS-derived neuron-astrocyte cocultures grown for the indicated times. N = 3 cell lines. (E) Representative images of MEA recordings from neuron-astrocyte cocultures at days 21, 30, 31, and 35. (F) Representative fluorescence images of indicated cell lines after immunocytochemical staining using antibodies against <t>ACE2.</t> Scale bar, 1,000 μm. (G) Mean normalized mRNA expression of ACE2 , TMPRSS2 , and NRP1 in Caco2, Caco2-ACE2, A549, neuron, and astrocyte cell lines. Values were normalized to human GAPDH expression. N = 2 to 4 cell lines.
    Ace2 Goat, supplied by Bio-Techne corporation, 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/result/ace2 goat/product/Bio-Techne corporation
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ace2 goat - by Bioz Stars, 2024-05
    96/100 stars

    Images

    1) Product Images from "SARS-CoV-2 Infection of Human Neurons Is TMPRSS2 Independent, Requires Endosomal Cell Entry, and Can Be Blocked by Inhibitors of Host Phosphoinositol-5 Kinase"

    Article Title: SARS-CoV-2 Infection of Human Neurons Is TMPRSS2 Independent, Requires Endosomal Cell Entry, and Can Be Blocked by Inhibitors of Host Phosphoinositol-5 Kinase

    Journal: Journal of Virology

    doi: 10.1128/jvi.00144-23

    Characterization of iPSC-derived neurons and astrocytes. (A) Representative images of immunocytochemical staining of iPSC-derived NGN2-neurons with MAP2, TUBB3, GABA, CUX1, and VGLUT1 antibodies. Scale bar, 50 μm. (B) Representative images of immunocytochemical staining of iPSC-derived astrocytes with GFAP, S100β, and AQP4 antibodies. Scale bar, 100 μm. (C) Mean normalized mRNA expression of astrocyte markers GFAP and S100β in iPSCs and iPSC-derived astrocytes. Values are normalized to human GAPDH expression. N = 4 cell lines. (D) Quantification of the mean firing rate (in hertz), percentage of electrodes partaking in bursts, percentage of electrodes partaking in network bursts, and network burst duration (in seconds) in iPCS-derived neuron-astrocyte cocultures grown for the indicated times. N = 3 cell lines. (E) Representative images of MEA recordings from neuron-astrocyte cocultures at days 21, 30, 31, and 35. (F) Representative fluorescence images of indicated cell lines after immunocytochemical staining using antibodies against ACE2. Scale bar, 1,000 μm. (G) Mean normalized mRNA expression of ACE2 , TMPRSS2 , and NRP1 in Caco2, Caco2-ACE2, A549, neuron, and astrocyte cell lines. Values were normalized to human GAPDH expression. N = 2 to 4 cell lines.
    Figure Legend Snippet: Characterization of iPSC-derived neurons and astrocytes. (A) Representative images of immunocytochemical staining of iPSC-derived NGN2-neurons with MAP2, TUBB3, GABA, CUX1, and VGLUT1 antibodies. Scale bar, 50 μm. (B) Representative images of immunocytochemical staining of iPSC-derived astrocytes with GFAP, S100β, and AQP4 antibodies. Scale bar, 100 μm. (C) Mean normalized mRNA expression of astrocyte markers GFAP and S100β in iPSCs and iPSC-derived astrocytes. Values are normalized to human GAPDH expression. N = 4 cell lines. (D) Quantification of the mean firing rate (in hertz), percentage of electrodes partaking in bursts, percentage of electrodes partaking in network bursts, and network burst duration (in seconds) in iPCS-derived neuron-astrocyte cocultures grown for the indicated times. N = 3 cell lines. (E) Representative images of MEA recordings from neuron-astrocyte cocultures at days 21, 30, 31, and 35. (F) Representative fluorescence images of indicated cell lines after immunocytochemical staining using antibodies against ACE2. Scale bar, 1,000 μm. (G) Mean normalized mRNA expression of ACE2 , TMPRSS2 , and NRP1 in Caco2, Caco2-ACE2, A549, neuron, and astrocyte cell lines. Values were normalized to human GAPDH expression. N = 2 to 4 cell lines.

    Techniques Used: Derivative Assay, Staining, Expressing, Fluorescence

    Infection of iPSC-derived neural cultures by SARS-CoV-2 is mainly dependent on the ACE2 receptor and does not spread efficiently. (A to C) Representative images of hiPCS-derived neuron-astrocyte cocultures infected with SARS-CoV-2 Wuhan virus for 48 h. Infected cells were identified by immunofluorescence detection (green) of the viral protein N (A and B) or N and double-stranded RNA colocalizing in the same infected cell (C). The image in panel B represents an enlarged area from the white boxed area in panel A. Nuclei were visualized with Hoechst DNA staining and neurons by immunodetection on MAP2 protein (magenta). (D) Percentage of SARS-COV-2-infected neurons in neuron-astrocyte cocultures at 24, 48, and 120 hpi. Neurons were identified by automated classification based on the fluorescence of MAP2. Astrocytes, identified as MAP2-negative cells, were never found infected. N = 6 wells per treatment. For each group, more than 10,000 cells were imaged and analyzed. (E) Representative fluorescence images of infected neuron-astrocyte cocultures from the experiment shown in panel D, at the indicated time points. SARS-CoV-2-infected cells (green, indicated by white arrows) identified by immunodetection of viral N protein, neurons by MAP2 (magenta), and nuclei (blue) by Hoechst DNA staining. (F) qRT-PCR of the extracellular medium withdrawn at the indicated time points after infection of neuron-astrocyte cocultures, using primers against SARS-COV-2 genome. Values are expressed as the number of genome copies per milliliter by comparison with a standard curve using known amounts of viral RNA. N = 6 wells per group. Significance was tested between 0 hpi and 24 hpi or 48 hpi (not significant [n.s.]). (G) Infection of Vero E6 cells with medium collected from infected neuron-astrocyte cocultures at 0, 24, or 48 hpi, or with the indicated amounts of a virus stock previously produced in Vero E6-TMPRSS2 cells (250 to 750 PFU or 25 to 75 PFU were used to infect each well of 96-well plates). N = 6 wells per group. Significance is shown in comparison to 0 hpi. (H) Representative immunofluorescent images of VERO E6 cells infected as described for panel G. Nuclei (blue) were stained with Hoechst, and infected cells (magenta) were identified by SARS-CoV-2 N protein immunofluorescence detection. (I) Percentage of SARS-COV-2-infected neurons in monocultures at 24 and 48 hpi. (J) Quantification of SARS-CoV-2 infection at 24 hpi in neurons that were treated with PBS (vehicle) or 2 μg/mL (low), 5 μg/mL (medium), or 20 μg/mL (high) anti-ACE2 or anti-actinin antibodies 1 h prior to infection. N = 6 wells per group. Significance is shown in comparison to the vehicle group. All scale bars are 100 μm. Columns and bars represent means ± SEM. Data were analyzed with an ordinary one-way ANOVA followed by Dunnett’s multiple-comparison test (C), an ordinary one-way ANOVA (D and E), or an ordinary one-way ANOVA followed by Tukey’s multiple-comparison test (J). *, P < 0.05; **, P < 0.005; ***, P < 0.0005.
    Figure Legend Snippet: Infection of iPSC-derived neural cultures by SARS-CoV-2 is mainly dependent on the ACE2 receptor and does not spread efficiently. (A to C) Representative images of hiPCS-derived neuron-astrocyte cocultures infected with SARS-CoV-2 Wuhan virus for 48 h. Infected cells were identified by immunofluorescence detection (green) of the viral protein N (A and B) or N and double-stranded RNA colocalizing in the same infected cell (C). The image in panel B represents an enlarged area from the white boxed area in panel A. Nuclei were visualized with Hoechst DNA staining and neurons by immunodetection on MAP2 protein (magenta). (D) Percentage of SARS-COV-2-infected neurons in neuron-astrocyte cocultures at 24, 48, and 120 hpi. Neurons were identified by automated classification based on the fluorescence of MAP2. Astrocytes, identified as MAP2-negative cells, were never found infected. N = 6 wells per treatment. For each group, more than 10,000 cells were imaged and analyzed. (E) Representative fluorescence images of infected neuron-astrocyte cocultures from the experiment shown in panel D, at the indicated time points. SARS-CoV-2-infected cells (green, indicated by white arrows) identified by immunodetection of viral N protein, neurons by MAP2 (magenta), and nuclei (blue) by Hoechst DNA staining. (F) qRT-PCR of the extracellular medium withdrawn at the indicated time points after infection of neuron-astrocyte cocultures, using primers against SARS-COV-2 genome. Values are expressed as the number of genome copies per milliliter by comparison with a standard curve using known amounts of viral RNA. N = 6 wells per group. Significance was tested between 0 hpi and 24 hpi or 48 hpi (not significant [n.s.]). (G) Infection of Vero E6 cells with medium collected from infected neuron-astrocyte cocultures at 0, 24, or 48 hpi, or with the indicated amounts of a virus stock previously produced in Vero E6-TMPRSS2 cells (250 to 750 PFU or 25 to 75 PFU were used to infect each well of 96-well plates). N = 6 wells per group. Significance is shown in comparison to 0 hpi. (H) Representative immunofluorescent images of VERO E6 cells infected as described for panel G. Nuclei (blue) were stained with Hoechst, and infected cells (magenta) were identified by SARS-CoV-2 N protein immunofluorescence detection. (I) Percentage of SARS-COV-2-infected neurons in monocultures at 24 and 48 hpi. (J) Quantification of SARS-CoV-2 infection at 24 hpi in neurons that were treated with PBS (vehicle) or 2 μg/mL (low), 5 μg/mL (medium), or 20 μg/mL (high) anti-ACE2 or anti-actinin antibodies 1 h prior to infection. N = 6 wells per group. Significance is shown in comparison to the vehicle group. All scale bars are 100 μm. Columns and bars represent means ± SEM. Data were analyzed with an ordinary one-way ANOVA followed by Dunnett’s multiple-comparison test (C), an ordinary one-way ANOVA (D and E), or an ordinary one-way ANOVA followed by Tukey’s multiple-comparison test (J). *, P < 0.05; **, P < 0.005; ***, P < 0.0005.

    Techniques Used: Infection, Derivative Assay, Virus, Immunofluorescence, Staining, Immunodetection, Fluorescence, Quantitative RT-PCR, Comparison, Produced

    SARS-CoV-2 infection of iPSC-derived neural cultures is blocked by inhibition of PIK5K but not serine proteases. (A and B) Treatment of neuron-astrocyte cocultures with 0.2 to 1 μM apilimod, 25 μM nafamostat, or a combination at 30 min prior to infection with SARS-CoV-2 at an MOI of 1.5. Analysis was at 24 hpi (A) and 48 hpi (B). N = 6 samples per group. (C) Treatment of Caco-2-ACE2 cells with 25 μM nafamostat, 1 μM apilimod, a combination, or anti-ACE2 antibody 30 min prior to infection with SARS-CoV-2 at an MOI of 2.5. Analysis was at 24 hpi. N = 3 samples per group. (D) Representative immunofluorescence images of SARS-CoV-2 infection (SARS-CoV-2 N protein) in Caco-2-ACE2 cells at 24 hpi at an MOI of 2.5, treated with the indicated drugs. Blue, Hoechst 33342; red, SARS-CoV-2 N. (E) Representative immunofluorescence images of SARS-CoV-2 infection (SARS-CoV-2 N protein) in neuron-astrocyte cocultures at 48 hpi at an MOI of 15, treated with the indicated drugs. Blue, Hoechst 33342; magenta, MAP2; green, SARS-CoV-2 N. (F) Treatment of neuron-astrocyte cocultures with 2 μM apilimod, 50 μM camostat, or a combination of the two drugs, 30 min prior to infection with SARS-CoV-2 at an equivalent MOI of 1.5. Cells were analyzed at 48 hpi after immunofluorescence imaging and image analysis. N = 3 wells per group. (G) Cytotoxicity assay in neuron-astrocyte cocultures treated for the indicated times with apilimod at concentrations ranging from 0.2 to 2 μM, or with 9 μM UCN-01. All scale bars are 100 μm. Columns and bars represent means ± SEM, respectively. Data were analyzed by an ordinary one-way ANOVA followed by Dunnett’s multiple-comparison test. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. Significance is shown compared to the vehicle group.
    Figure Legend Snippet: SARS-CoV-2 infection of iPSC-derived neural cultures is blocked by inhibition of PIK5K but not serine proteases. (A and B) Treatment of neuron-astrocyte cocultures with 0.2 to 1 μM apilimod, 25 μM nafamostat, or a combination at 30 min prior to infection with SARS-CoV-2 at an MOI of 1.5. Analysis was at 24 hpi (A) and 48 hpi (B). N = 6 samples per group. (C) Treatment of Caco-2-ACE2 cells with 25 μM nafamostat, 1 μM apilimod, a combination, or anti-ACE2 antibody 30 min prior to infection with SARS-CoV-2 at an MOI of 2.5. Analysis was at 24 hpi. N = 3 samples per group. (D) Representative immunofluorescence images of SARS-CoV-2 infection (SARS-CoV-2 N protein) in Caco-2-ACE2 cells at 24 hpi at an MOI of 2.5, treated with the indicated drugs. Blue, Hoechst 33342; red, SARS-CoV-2 N. (E) Representative immunofluorescence images of SARS-CoV-2 infection (SARS-CoV-2 N protein) in neuron-astrocyte cocultures at 48 hpi at an MOI of 15, treated with the indicated drugs. Blue, Hoechst 33342; magenta, MAP2; green, SARS-CoV-2 N. (F) Treatment of neuron-astrocyte cocultures with 2 μM apilimod, 50 μM camostat, or a combination of the two drugs, 30 min prior to infection with SARS-CoV-2 at an equivalent MOI of 1.5. Cells were analyzed at 48 hpi after immunofluorescence imaging and image analysis. N = 3 wells per group. (G) Cytotoxicity assay in neuron-astrocyte cocultures treated for the indicated times with apilimod at concentrations ranging from 0.2 to 2 μM, or with 9 μM UCN-01. All scale bars are 100 μm. Columns and bars represent means ± SEM, respectively. Data were analyzed by an ordinary one-way ANOVA followed by Dunnett’s multiple-comparison test. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. Significance is shown compared to the vehicle group.

    Techniques Used: Infection, Derivative Assay, Inhibition, Immunofluorescence, Imaging, Cytotoxicity Assay, Comparison

    Cathepsin L inhibitor SB412515 does not significantly inhibit SARS-CoV-2 neuron infection. (A) Quantification of SARS-CoV-2 infection in neuron-astrocyte cocultures (48 hpi), Vero E6, A549-ACE2-TMRPSS2, and Caco2-ACE2 cells (24 hpi) treated with 10 μM cathepsin L inhibitor SB412515 starting 30 min before infection. (B) Representative immunofluorescence images of cells infected and treated as described for panel A. Blue, nuclei stained with Hoechst; magenta, infected cells; yellow, neurons (identified by immunofluorescence detection of SARS-VOV-2 N and MAP2 proteins, respectively). Columns and bars represent means ± SEM, respectively. Data were analyzed by an ordinary one-way ANOVA followed by Dunnett’s multiple-comparison test. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. Significance is shown compared to the vehicle group.
    Figure Legend Snippet: Cathepsin L inhibitor SB412515 does not significantly inhibit SARS-CoV-2 neuron infection. (A) Quantification of SARS-CoV-2 infection in neuron-astrocyte cocultures (48 hpi), Vero E6, A549-ACE2-TMRPSS2, and Caco2-ACE2 cells (24 hpi) treated with 10 μM cathepsin L inhibitor SB412515 starting 30 min before infection. (B) Representative immunofluorescence images of cells infected and treated as described for panel A. Blue, nuclei stained with Hoechst; magenta, infected cells; yellow, neurons (identified by immunofluorescence detection of SARS-VOV-2 N and MAP2 proteins, respectively). Columns and bars represent means ± SEM, respectively. Data were analyzed by an ordinary one-way ANOVA followed by Dunnett’s multiple-comparison test. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. Significance is shown compared to the vehicle group.

    Techniques Used: Infection, Immunofluorescence, Staining, Comparison

    Antibodies used in the study
    Figure Legend Snippet: Antibodies used in the study

    Techniques Used:

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96
    Bio-Techne corporation hace2
    Hace2, supplied by Bio-Techne corporation, 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/result/hace2/product/Bio-Techne corporation
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hace2 - by Bioz Stars, 2024-05
    96/100 stars
      Buy from Supplier

    96
    Bio-Techne corporation anti ace2
    ( A ) (Top) Mean (+/− SEM) of SFV viral titers in NoDice FHA:DICER WT and N1 cells infected at an MOI of 1.10 −4 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Unpaired t-test with Welch’s correction. * p < 0.05. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in SFV infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( B ) (Top) Mean (+/− SEM) of EV71 viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 0.1 for 24 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test. ** p < 0.01. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in EV71 infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( C ) (Top) Mean (+/− SEM) of VSV-GFP viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 1.10 −5 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Unpaired t-test. ns: non-significant. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in VSV-GFP infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( D ) Western blot analysis of human <t>ACE2</t> and HA (for DICER) expression in NoDice FHA:DICER WT and N1 cells ( n = 3 biological replicates). Alpha-Tubulin was used as loading control. ( E ) Western blot analysis of DICER and NUCLEOCAPSID expression in SARS-CoV-2 infected NoDice FHA:DICER WT and N1 cells at an MOI of 0.001 for 48 h. GAPDH was used as loading control. Band intensity was quantified and normalized to GAPDH for three independent biological replicates, then represented as mean (+/− SD) under the corresponding lane. ( F ) Mean (+/− SEM) of SARS-CoV-2 viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 0.001 for 48 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test. * p < 0.05. .
    Anti Ace2, supplied by Bio-Techne corporation, 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/result/anti ace2/product/Bio-Techne corporation
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti ace2 - by Bioz Stars, 2024-05
    96/100 stars
      Buy from Supplier

    94
    Bio-Techne corporation ace2 antibody
    ( A ) (Top) Mean (+/− SEM) of SFV viral titers in NoDice FHA:DICER WT and N1 cells infected at an MOI of 1.10 −4 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Unpaired t-test with Welch’s correction. * p < 0.05. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in SFV infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( B ) (Top) Mean (+/− SEM) of EV71 viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 0.1 for 24 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test. ** p < 0.01. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in EV71 infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( C ) (Top) Mean (+/− SEM) of VSV-GFP viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 1.10 −5 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Unpaired t-test. ns: non-significant. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in VSV-GFP infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( D ) Western blot analysis of human <t>ACE2</t> and HA (for DICER) expression in NoDice FHA:DICER WT and N1 cells ( n = 3 biological replicates). Alpha-Tubulin was used as loading control. ( E ) Western blot analysis of DICER and NUCLEOCAPSID expression in SARS-CoV-2 infected NoDice FHA:DICER WT and N1 cells at an MOI of 0.001 for 48 h. GAPDH was used as loading control. Band intensity was quantified and normalized to GAPDH for three independent biological replicates, then represented as mean (+/− SD) under the corresponding lane. ( F ) Mean (+/− SEM) of SARS-CoV-2 viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 0.001 for 48 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test. * p < 0.05. .
    Ace2 Antibody, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ace2 antibody/product/Bio-Techne corporation
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ace2 antibody - by Bioz Stars, 2024-05
    94/100 stars
      Buy from Supplier

    93
    Bio-Techne corporation antibodies to ace2
    ( A ) (Top) Mean (+/− SEM) of SFV viral titers in NoDice FHA:DICER WT and N1 cells infected at an MOI of 1.10 −4 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Unpaired t-test with Welch’s correction. * p < 0.05. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in SFV infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( B ) (Top) Mean (+/− SEM) of EV71 viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 0.1 for 24 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test. ** p < 0.01. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in EV71 infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( C ) (Top) Mean (+/− SEM) of VSV-GFP viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 1.10 −5 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Unpaired t-test. ns: non-significant. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in VSV-GFP infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( D ) Western blot analysis of human <t>ACE2</t> and HA (for DICER) expression in NoDice FHA:DICER WT and N1 cells ( n = 3 biological replicates). Alpha-Tubulin was used as loading control. ( E ) Western blot analysis of DICER and NUCLEOCAPSID expression in SARS-CoV-2 infected NoDice FHA:DICER WT and N1 cells at an MOI of 0.001 for 48 h. GAPDH was used as loading control. Band intensity was quantified and normalized to GAPDH for three independent biological replicates, then represented as mean (+/− SD) under the corresponding lane. ( F ) Mean (+/− SEM) of SARS-CoV-2 viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 0.001 for 48 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test. * p < 0.05. .
    Antibodies To Ace2, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/antibodies to ace2/product/Bio-Techne corporation
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    antibodies to ace2 - by Bioz Stars, 2024-05
    93/100 stars
      Buy from Supplier

    96
    Bio-Techne corporation goat anti ace2
    ( A ) (Top) Mean (+/− SEM) of SFV viral titers in NoDice FHA:DICER WT and N1 cells infected at an MOI of 1.10 −4 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Unpaired t-test with Welch’s correction. * p < 0.05. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in SFV infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( B ) (Top) Mean (+/− SEM) of EV71 viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 0.1 for 24 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test. ** p < 0.01. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in EV71 infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( C ) (Top) Mean (+/− SEM) of VSV-GFP viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 1.10 −5 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Unpaired t-test. ns: non-significant. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in VSV-GFP infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( D ) Western blot analysis of human <t>ACE2</t> and HA (for DICER) expression in NoDice FHA:DICER WT and N1 cells ( n = 3 biological replicates). Alpha-Tubulin was used as loading control. ( E ) Western blot analysis of DICER and NUCLEOCAPSID expression in SARS-CoV-2 infected NoDice FHA:DICER WT and N1 cells at an MOI of 0.001 for 48 h. GAPDH was used as loading control. Band intensity was quantified and normalized to GAPDH for three independent biological replicates, then represented as mean (+/− SD) under the corresponding lane. ( F ) Mean (+/− SEM) of SARS-CoV-2 viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 0.001 for 48 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test. * p < 0.05. .
    Goat Anti Ace2, supplied by Bio-Techne corporation, 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/result/goat anti ace2/product/Bio-Techne corporation
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti ace2 - by Bioz Stars, 2024-05
    96/100 stars
      Buy from Supplier

    96
    Bio-Techne corporation ace2 goat
    Characterization of iPSC-derived neurons and astrocytes. (A) Representative images of immunocytochemical staining of iPSC-derived NGN2-neurons with MAP2, TUBB3, GABA, CUX1, and VGLUT1 antibodies. Scale bar, 50 μm. (B) Representative images of immunocytochemical staining of iPSC-derived astrocytes with GFAP, S100β, and AQP4 antibodies. Scale bar, 100 μm. (C) Mean normalized mRNA expression of astrocyte markers GFAP and S100β in iPSCs and iPSC-derived astrocytes. Values are normalized to human GAPDH expression. N = 4 cell lines. (D) Quantification of the mean firing rate (in hertz), percentage of electrodes partaking in bursts, percentage of electrodes partaking in network bursts, and network burst duration (in seconds) in iPCS-derived neuron-astrocyte cocultures grown for the indicated times. N = 3 cell lines. (E) Representative images of MEA recordings from neuron-astrocyte cocultures at days 21, 30, 31, and 35. (F) Representative fluorescence images of indicated cell lines after immunocytochemical staining using antibodies against <t>ACE2.</t> Scale bar, 1,000 μm. (G) Mean normalized mRNA expression of ACE2 , TMPRSS2 , and NRP1 in Caco2, Caco2-ACE2, A549, neuron, and astrocyte cell lines. Values were normalized to human GAPDH expression. N = 2 to 4 cell lines.
    Ace2 Goat, supplied by Bio-Techne corporation, 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/result/ace2 goat/product/Bio-Techne corporation
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ace2 goat - by Bioz Stars, 2024-05
    96/100 stars
      Buy from Supplier

    Image Search Results


    ( A ) (Top) Mean (+/− SEM) of SFV viral titers in NoDice FHA:DICER WT and N1 cells infected at an MOI of 1.10 −4 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Unpaired t-test with Welch’s correction. * p < 0.05. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in SFV infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( B ) (Top) Mean (+/− SEM) of EV71 viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 0.1 for 24 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test. ** p < 0.01. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in EV71 infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( C ) (Top) Mean (+/− SEM) of VSV-GFP viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 1.10 −5 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Unpaired t-test. ns: non-significant. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in VSV-GFP infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( D ) Western blot analysis of human ACE2 and HA (for DICER) expression in NoDice FHA:DICER WT and N1 cells ( n = 3 biological replicates). Alpha-Tubulin was used as loading control. ( E ) Western blot analysis of DICER and NUCLEOCAPSID expression in SARS-CoV-2 infected NoDice FHA:DICER WT and N1 cells at an MOI of 0.001 for 48 h. GAPDH was used as loading control. Band intensity was quantified and normalized to GAPDH for three independent biological replicates, then represented as mean (+/− SD) under the corresponding lane. ( F ) Mean (+/− SEM) of SARS-CoV-2 viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 0.001 for 48 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test. * p < 0.05. .

    Journal: The EMBO Journal

    Article Title: The helicase domain of human Dicer prevents RNAi-independent activation of antiviral and inflammatory pathways

    doi: 10.1038/s44318-024-00035-2

    Figure Lengend Snippet: ( A ) (Top) Mean (+/− SEM) of SFV viral titers in NoDice FHA:DICER WT and N1 cells infected at an MOI of 1.10 −4 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Unpaired t-test with Welch’s correction. * p < 0.05. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in SFV infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( B ) (Top) Mean (+/− SEM) of EV71 viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 0.1 for 24 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test. ** p < 0.01. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in EV71 infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( C ) (Top) Mean (+/− SEM) of VSV-GFP viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 1.10 −5 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Unpaired t-test. ns: non-significant. (Bottom) Immunofluorescence analysis on NoDice FHA:DICER WT and N1 cells in VSV-GFP infected cells. J2 antibody (red) was used to detect dsRNA upon infection. DAPI was used to stain the nuclei (blue). Scale bar = 100 µm, n = 3 biological replicates. ( D ) Western blot analysis of human ACE2 and HA (for DICER) expression in NoDice FHA:DICER WT and N1 cells ( n = 3 biological replicates). Alpha-Tubulin was used as loading control. ( E ) Western blot analysis of DICER and NUCLEOCAPSID expression in SARS-CoV-2 infected NoDice FHA:DICER WT and N1 cells at an MOI of 0.001 for 48 h. GAPDH was used as loading control. Band intensity was quantified and normalized to GAPDH for three independent biological replicates, then represented as mean (+/− SD) under the corresponding lane. ( F ) Mean (+/− SEM) of SARS-CoV-2 viral titers in NoDice FHA:DICER WT and N1 infected at an MOI of 0.001 for 48 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test. * p < 0.05. .

    Article Snippet: Gunter Meister, University of Regensburg), anti-ACE2 (1:1000, AF933 Bio-Techne), anti-NF-kB/p65 (1:1000, #8242 Cell Signaling), anti-p-NF-kB/p65 (1:1000, #3033 Cell Signaling) and anti-IkBα (1:1000, #4812 Cell Signaling).

    Techniques: Infection, Plaque Assay, Immunofluorescence, Staining, Western Blot, Expressing

    ( A ) Schematic representation of PKR and its two point mutants: K296R and T451A. ( B ) Western blot analysis of DICER, p-PKR, PKR, and CAPSID expression in SINV-GFP infected NoDice∆PKR FHA:DICER WT cells expressing MYC:EMPTY CTRL vector, MYC:PKR, MYC K296R or MYC:T451A at an MOI of 0.02 for 24 h. Alpha-Tubulin was used as loading control. Band intensity was quantified and normalized to Tubulin for three independent biological replicates, then represented as mean (+/− SD) under the corresponding lane; p-PKR = p-PKR/PKR quantification. ( C ) Mean (+/− SEM) of SINV-GFP viral titers in the same samples as in (B), infected at an MOI of 0.02 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Ordinary one-way ANOVA test with Dunnett’s correction. ns: non-significant. ( D ) Western blot analysis of DICER, p-PKR, PKR and CAPSID expression in SINV-GFP infected NoDice∆PKR FHA:DICER N1 cells expressing MYC:EMPTY CTRL vector, MYC:PKR, MYC K296R or MYC:T451A at an MOI of 0.02 for 24 h. Alpha-Tubulin was used as loading control. Band intensity was quantified and normalized to Tubulin for three independent biological replicates, then represented as mean (+/− SD) under the corresponding lane; p-PKR = p-PKR/PKR quantification. ( E ) Mean (+/− SEM) of SINV-GFP viral titers in the same samples as in ( D ), infected at an MOI of 0.02 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Ordinary one-way ANOVA test with Dunnett’s correction. * p < 0.05. ( F ) Western blot analysis of DICER, human ACE2, PKR and NUCLEOCAPSID expression in SARS-CoV-2 infected NoDice FHA:DICER WT or N1 and NoDice∆PKR FHA:DICER WT or N1 cells at an MOI of 0.001 for 48 h. GAPDH was used as loading control. Band intensity was quantified and normalized to GAPDH for three independent biological replicates, then represented as mean (+/− SD) under the corresponding lane. ( G ) Mean (+/− SEM) of SARS-CoV-2 viral titers in the same samples as in ( F ), infected at an MOI of 0.001 for 48 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test with Welch’s correction. * p < 0.05; ** p < 0.01. .

    Journal: The EMBO Journal

    Article Title: The helicase domain of human Dicer prevents RNAi-independent activation of antiviral and inflammatory pathways

    doi: 10.1038/s44318-024-00035-2

    Figure Lengend Snippet: ( A ) Schematic representation of PKR and its two point mutants: K296R and T451A. ( B ) Western blot analysis of DICER, p-PKR, PKR, and CAPSID expression in SINV-GFP infected NoDice∆PKR FHA:DICER WT cells expressing MYC:EMPTY CTRL vector, MYC:PKR, MYC K296R or MYC:T451A at an MOI of 0.02 for 24 h. Alpha-Tubulin was used as loading control. Band intensity was quantified and normalized to Tubulin for three independent biological replicates, then represented as mean (+/− SD) under the corresponding lane; p-PKR = p-PKR/PKR quantification. ( C ) Mean (+/− SEM) of SINV-GFP viral titers in the same samples as in (B), infected at an MOI of 0.02 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Ordinary one-way ANOVA test with Dunnett’s correction. ns: non-significant. ( D ) Western blot analysis of DICER, p-PKR, PKR and CAPSID expression in SINV-GFP infected NoDice∆PKR FHA:DICER N1 cells expressing MYC:EMPTY CTRL vector, MYC:PKR, MYC K296R or MYC:T451A at an MOI of 0.02 for 24 h. Alpha-Tubulin was used as loading control. Band intensity was quantified and normalized to Tubulin for three independent biological replicates, then represented as mean (+/− SD) under the corresponding lane; p-PKR = p-PKR/PKR quantification. ( E ) Mean (+/− SEM) of SINV-GFP viral titers in the same samples as in ( D ), infected at an MOI of 0.02 for 24 h ( n = 3 biological replicates) from plaque assay quantification. Ordinary one-way ANOVA test with Dunnett’s correction. * p < 0.05. ( F ) Western blot analysis of DICER, human ACE2, PKR and NUCLEOCAPSID expression in SARS-CoV-2 infected NoDice FHA:DICER WT or N1 and NoDice∆PKR FHA:DICER WT or N1 cells at an MOI of 0.001 for 48 h. GAPDH was used as loading control. Band intensity was quantified and normalized to GAPDH for three independent biological replicates, then represented as mean (+/− SD) under the corresponding lane. ( G ) Mean (+/− SEM) of SARS-CoV-2 viral titers in the same samples as in ( F ), infected at an MOI of 0.001 for 48 h ( n = 3 biological replicates) from TCID50 quantification. Unpaired t-test with Welch’s correction. * p < 0.05; ** p < 0.01. .

    Article Snippet: Gunter Meister, University of Regensburg), anti-ACE2 (1:1000, AF933 Bio-Techne), anti-NF-kB/p65 (1:1000, #8242 Cell Signaling), anti-p-NF-kB/p65 (1:1000, #3033 Cell Signaling) and anti-IkBα (1:1000, #4812 Cell Signaling).

    Techniques: Western Blot, Expressing, Infection, Plasmid Preparation, Plaque Assay

    Characterization of iPSC-derived neurons and astrocytes. (A) Representative images of immunocytochemical staining of iPSC-derived NGN2-neurons with MAP2, TUBB3, GABA, CUX1, and VGLUT1 antibodies. Scale bar, 50 μm. (B) Representative images of immunocytochemical staining of iPSC-derived astrocytes with GFAP, S100β, and AQP4 antibodies. Scale bar, 100 μm. (C) Mean normalized mRNA expression of astrocyte markers GFAP and S100β in iPSCs and iPSC-derived astrocytes. Values are normalized to human GAPDH expression. N = 4 cell lines. (D) Quantification of the mean firing rate (in hertz), percentage of electrodes partaking in bursts, percentage of electrodes partaking in network bursts, and network burst duration (in seconds) in iPCS-derived neuron-astrocyte cocultures grown for the indicated times. N = 3 cell lines. (E) Representative images of MEA recordings from neuron-astrocyte cocultures at days 21, 30, 31, and 35. (F) Representative fluorescence images of indicated cell lines after immunocytochemical staining using antibodies against ACE2. Scale bar, 1,000 μm. (G) Mean normalized mRNA expression of ACE2 , TMPRSS2 , and NRP1 in Caco2, Caco2-ACE2, A549, neuron, and astrocyte cell lines. Values were normalized to human GAPDH expression. N = 2 to 4 cell lines.

    Journal: Journal of Virology

    Article Title: SARS-CoV-2 Infection of Human Neurons Is TMPRSS2 Independent, Requires Endosomal Cell Entry, and Can Be Blocked by Inhibitors of Host Phosphoinositol-5 Kinase

    doi: 10.1128/jvi.00144-23

    Figure Lengend Snippet: Characterization of iPSC-derived neurons and astrocytes. (A) Representative images of immunocytochemical staining of iPSC-derived NGN2-neurons with MAP2, TUBB3, GABA, CUX1, and VGLUT1 antibodies. Scale bar, 50 μm. (B) Representative images of immunocytochemical staining of iPSC-derived astrocytes with GFAP, S100β, and AQP4 antibodies. Scale bar, 100 μm. (C) Mean normalized mRNA expression of astrocyte markers GFAP and S100β in iPSCs and iPSC-derived astrocytes. Values are normalized to human GAPDH expression. N = 4 cell lines. (D) Quantification of the mean firing rate (in hertz), percentage of electrodes partaking in bursts, percentage of electrodes partaking in network bursts, and network burst duration (in seconds) in iPCS-derived neuron-astrocyte cocultures grown for the indicated times. N = 3 cell lines. (E) Representative images of MEA recordings from neuron-astrocyte cocultures at days 21, 30, 31, and 35. (F) Representative fluorescence images of indicated cell lines after immunocytochemical staining using antibodies against ACE2. Scale bar, 1,000 μm. (G) Mean normalized mRNA expression of ACE2 , TMPRSS2 , and NRP1 in Caco2, Caco2-ACE2, A549, neuron, and astrocyte cell lines. Values were normalized to human GAPDH expression. N = 2 to 4 cell lines.

    Article Snippet: ACE2 goat , 1:1,000 , Bio-Techne , AF933.

    Techniques: Derivative Assay, Staining, Expressing, Fluorescence

    Infection of iPSC-derived neural cultures by SARS-CoV-2 is mainly dependent on the ACE2 receptor and does not spread efficiently. (A to C) Representative images of hiPCS-derived neuron-astrocyte cocultures infected with SARS-CoV-2 Wuhan virus for 48 h. Infected cells were identified by immunofluorescence detection (green) of the viral protein N (A and B) or N and double-stranded RNA colocalizing in the same infected cell (C). The image in panel B represents an enlarged area from the white boxed area in panel A. Nuclei were visualized with Hoechst DNA staining and neurons by immunodetection on MAP2 protein (magenta). (D) Percentage of SARS-COV-2-infected neurons in neuron-astrocyte cocultures at 24, 48, and 120 hpi. Neurons were identified by automated classification based on the fluorescence of MAP2. Astrocytes, identified as MAP2-negative cells, were never found infected. N = 6 wells per treatment. For each group, more than 10,000 cells were imaged and analyzed. (E) Representative fluorescence images of infected neuron-astrocyte cocultures from the experiment shown in panel D, at the indicated time points. SARS-CoV-2-infected cells (green, indicated by white arrows) identified by immunodetection of viral N protein, neurons by MAP2 (magenta), and nuclei (blue) by Hoechst DNA staining. (F) qRT-PCR of the extracellular medium withdrawn at the indicated time points after infection of neuron-astrocyte cocultures, using primers against SARS-COV-2 genome. Values are expressed as the number of genome copies per milliliter by comparison with a standard curve using known amounts of viral RNA. N = 6 wells per group. Significance was tested between 0 hpi and 24 hpi or 48 hpi (not significant [n.s.]). (G) Infection of Vero E6 cells with medium collected from infected neuron-astrocyte cocultures at 0, 24, or 48 hpi, or with the indicated amounts of a virus stock previously produced in Vero E6-TMPRSS2 cells (250 to 750 PFU or 25 to 75 PFU were used to infect each well of 96-well plates). N = 6 wells per group. Significance is shown in comparison to 0 hpi. (H) Representative immunofluorescent images of VERO E6 cells infected as described for panel G. Nuclei (blue) were stained with Hoechst, and infected cells (magenta) were identified by SARS-CoV-2 N protein immunofluorescence detection. (I) Percentage of SARS-COV-2-infected neurons in monocultures at 24 and 48 hpi. (J) Quantification of SARS-CoV-2 infection at 24 hpi in neurons that were treated with PBS (vehicle) or 2 μg/mL (low), 5 μg/mL (medium), or 20 μg/mL (high) anti-ACE2 or anti-actinin antibodies 1 h prior to infection. N = 6 wells per group. Significance is shown in comparison to the vehicle group. All scale bars are 100 μm. Columns and bars represent means ± SEM. Data were analyzed with an ordinary one-way ANOVA followed by Dunnett’s multiple-comparison test (C), an ordinary one-way ANOVA (D and E), or an ordinary one-way ANOVA followed by Tukey’s multiple-comparison test (J). *, P < 0.05; **, P < 0.005; ***, P < 0.0005.

    Journal: Journal of Virology

    Article Title: SARS-CoV-2 Infection of Human Neurons Is TMPRSS2 Independent, Requires Endosomal Cell Entry, and Can Be Blocked by Inhibitors of Host Phosphoinositol-5 Kinase

    doi: 10.1128/jvi.00144-23

    Figure Lengend Snippet: Infection of iPSC-derived neural cultures by SARS-CoV-2 is mainly dependent on the ACE2 receptor and does not spread efficiently. (A to C) Representative images of hiPCS-derived neuron-astrocyte cocultures infected with SARS-CoV-2 Wuhan virus for 48 h. Infected cells were identified by immunofluorescence detection (green) of the viral protein N (A and B) or N and double-stranded RNA colocalizing in the same infected cell (C). The image in panel B represents an enlarged area from the white boxed area in panel A. Nuclei were visualized with Hoechst DNA staining and neurons by immunodetection on MAP2 protein (magenta). (D) Percentage of SARS-COV-2-infected neurons in neuron-astrocyte cocultures at 24, 48, and 120 hpi. Neurons were identified by automated classification based on the fluorescence of MAP2. Astrocytes, identified as MAP2-negative cells, were never found infected. N = 6 wells per treatment. For each group, more than 10,000 cells were imaged and analyzed. (E) Representative fluorescence images of infected neuron-astrocyte cocultures from the experiment shown in panel D, at the indicated time points. SARS-CoV-2-infected cells (green, indicated by white arrows) identified by immunodetection of viral N protein, neurons by MAP2 (magenta), and nuclei (blue) by Hoechst DNA staining. (F) qRT-PCR of the extracellular medium withdrawn at the indicated time points after infection of neuron-astrocyte cocultures, using primers against SARS-COV-2 genome. Values are expressed as the number of genome copies per milliliter by comparison with a standard curve using known amounts of viral RNA. N = 6 wells per group. Significance was tested between 0 hpi and 24 hpi or 48 hpi (not significant [n.s.]). (G) Infection of Vero E6 cells with medium collected from infected neuron-astrocyte cocultures at 0, 24, or 48 hpi, or with the indicated amounts of a virus stock previously produced in Vero E6-TMPRSS2 cells (250 to 750 PFU or 25 to 75 PFU were used to infect each well of 96-well plates). N = 6 wells per group. Significance is shown in comparison to 0 hpi. (H) Representative immunofluorescent images of VERO E6 cells infected as described for panel G. Nuclei (blue) were stained with Hoechst, and infected cells (magenta) were identified by SARS-CoV-2 N protein immunofluorescence detection. (I) Percentage of SARS-COV-2-infected neurons in monocultures at 24 and 48 hpi. (J) Quantification of SARS-CoV-2 infection at 24 hpi in neurons that were treated with PBS (vehicle) or 2 μg/mL (low), 5 μg/mL (medium), or 20 μg/mL (high) anti-ACE2 or anti-actinin antibodies 1 h prior to infection. N = 6 wells per group. Significance is shown in comparison to the vehicle group. All scale bars are 100 μm. Columns and bars represent means ± SEM. Data were analyzed with an ordinary one-way ANOVA followed by Dunnett’s multiple-comparison test (C), an ordinary one-way ANOVA (D and E), or an ordinary one-way ANOVA followed by Tukey’s multiple-comparison test (J). *, P < 0.05; **, P < 0.005; ***, P < 0.0005.

    Article Snippet: ACE2 goat , 1:1,000 , Bio-Techne , AF933.

    Techniques: Infection, Derivative Assay, Virus, Immunofluorescence, Staining, Immunodetection, Fluorescence, Quantitative RT-PCR, Comparison, Produced

    SARS-CoV-2 infection of iPSC-derived neural cultures is blocked by inhibition of PIK5K but not serine proteases. (A and B) Treatment of neuron-astrocyte cocultures with 0.2 to 1 μM apilimod, 25 μM nafamostat, or a combination at 30 min prior to infection with SARS-CoV-2 at an MOI of 1.5. Analysis was at 24 hpi (A) and 48 hpi (B). N = 6 samples per group. (C) Treatment of Caco-2-ACE2 cells with 25 μM nafamostat, 1 μM apilimod, a combination, or anti-ACE2 antibody 30 min prior to infection with SARS-CoV-2 at an MOI of 2.5. Analysis was at 24 hpi. N = 3 samples per group. (D) Representative immunofluorescence images of SARS-CoV-2 infection (SARS-CoV-2 N protein) in Caco-2-ACE2 cells at 24 hpi at an MOI of 2.5, treated with the indicated drugs. Blue, Hoechst 33342; red, SARS-CoV-2 N. (E) Representative immunofluorescence images of SARS-CoV-2 infection (SARS-CoV-2 N protein) in neuron-astrocyte cocultures at 48 hpi at an MOI of 15, treated with the indicated drugs. Blue, Hoechst 33342; magenta, MAP2; green, SARS-CoV-2 N. (F) Treatment of neuron-astrocyte cocultures with 2 μM apilimod, 50 μM camostat, or a combination of the two drugs, 30 min prior to infection with SARS-CoV-2 at an equivalent MOI of 1.5. Cells were analyzed at 48 hpi after immunofluorescence imaging and image analysis. N = 3 wells per group. (G) Cytotoxicity assay in neuron-astrocyte cocultures treated for the indicated times with apilimod at concentrations ranging from 0.2 to 2 μM, or with 9 μM UCN-01. All scale bars are 100 μm. Columns and bars represent means ± SEM, respectively. Data were analyzed by an ordinary one-way ANOVA followed by Dunnett’s multiple-comparison test. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. Significance is shown compared to the vehicle group.

    Journal: Journal of Virology

    Article Title: SARS-CoV-2 Infection of Human Neurons Is TMPRSS2 Independent, Requires Endosomal Cell Entry, and Can Be Blocked by Inhibitors of Host Phosphoinositol-5 Kinase

    doi: 10.1128/jvi.00144-23

    Figure Lengend Snippet: SARS-CoV-2 infection of iPSC-derived neural cultures is blocked by inhibition of PIK5K but not serine proteases. (A and B) Treatment of neuron-astrocyte cocultures with 0.2 to 1 μM apilimod, 25 μM nafamostat, or a combination at 30 min prior to infection with SARS-CoV-2 at an MOI of 1.5. Analysis was at 24 hpi (A) and 48 hpi (B). N = 6 samples per group. (C) Treatment of Caco-2-ACE2 cells with 25 μM nafamostat, 1 μM apilimod, a combination, or anti-ACE2 antibody 30 min prior to infection with SARS-CoV-2 at an MOI of 2.5. Analysis was at 24 hpi. N = 3 samples per group. (D) Representative immunofluorescence images of SARS-CoV-2 infection (SARS-CoV-2 N protein) in Caco-2-ACE2 cells at 24 hpi at an MOI of 2.5, treated with the indicated drugs. Blue, Hoechst 33342; red, SARS-CoV-2 N. (E) Representative immunofluorescence images of SARS-CoV-2 infection (SARS-CoV-2 N protein) in neuron-astrocyte cocultures at 48 hpi at an MOI of 15, treated with the indicated drugs. Blue, Hoechst 33342; magenta, MAP2; green, SARS-CoV-2 N. (F) Treatment of neuron-astrocyte cocultures with 2 μM apilimod, 50 μM camostat, or a combination of the two drugs, 30 min prior to infection with SARS-CoV-2 at an equivalent MOI of 1.5. Cells were analyzed at 48 hpi after immunofluorescence imaging and image analysis. N = 3 wells per group. (G) Cytotoxicity assay in neuron-astrocyte cocultures treated for the indicated times with apilimod at concentrations ranging from 0.2 to 2 μM, or with 9 μM UCN-01. All scale bars are 100 μm. Columns and bars represent means ± SEM, respectively. Data were analyzed by an ordinary one-way ANOVA followed by Dunnett’s multiple-comparison test. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. Significance is shown compared to the vehicle group.

    Article Snippet: ACE2 goat , 1:1,000 , Bio-Techne , AF933.

    Techniques: Infection, Derivative Assay, Inhibition, Immunofluorescence, Imaging, Cytotoxicity Assay, Comparison

    Cathepsin L inhibitor SB412515 does not significantly inhibit SARS-CoV-2 neuron infection. (A) Quantification of SARS-CoV-2 infection in neuron-astrocyte cocultures (48 hpi), Vero E6, A549-ACE2-TMRPSS2, and Caco2-ACE2 cells (24 hpi) treated with 10 μM cathepsin L inhibitor SB412515 starting 30 min before infection. (B) Representative immunofluorescence images of cells infected and treated as described for panel A. Blue, nuclei stained with Hoechst; magenta, infected cells; yellow, neurons (identified by immunofluorescence detection of SARS-VOV-2 N and MAP2 proteins, respectively). Columns and bars represent means ± SEM, respectively. Data were analyzed by an ordinary one-way ANOVA followed by Dunnett’s multiple-comparison test. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. Significance is shown compared to the vehicle group.

    Journal: Journal of Virology

    Article Title: SARS-CoV-2 Infection of Human Neurons Is TMPRSS2 Independent, Requires Endosomal Cell Entry, and Can Be Blocked by Inhibitors of Host Phosphoinositol-5 Kinase

    doi: 10.1128/jvi.00144-23

    Figure Lengend Snippet: Cathepsin L inhibitor SB412515 does not significantly inhibit SARS-CoV-2 neuron infection. (A) Quantification of SARS-CoV-2 infection in neuron-astrocyte cocultures (48 hpi), Vero E6, A549-ACE2-TMRPSS2, and Caco2-ACE2 cells (24 hpi) treated with 10 μM cathepsin L inhibitor SB412515 starting 30 min before infection. (B) Representative immunofluorescence images of cells infected and treated as described for panel A. Blue, nuclei stained with Hoechst; magenta, infected cells; yellow, neurons (identified by immunofluorescence detection of SARS-VOV-2 N and MAP2 proteins, respectively). Columns and bars represent means ± SEM, respectively. Data were analyzed by an ordinary one-way ANOVA followed by Dunnett’s multiple-comparison test. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. Significance is shown compared to the vehicle group.

    Article Snippet: ACE2 goat , 1:1,000 , Bio-Techne , AF933.

    Techniques: Infection, Immunofluorescence, Staining, Comparison

    Antibodies used in the study

    Journal: Journal of Virology

    Article Title: SARS-CoV-2 Infection of Human Neurons Is TMPRSS2 Independent, Requires Endosomal Cell Entry, and Can Be Blocked by Inhibitors of Host Phosphoinositol-5 Kinase

    doi: 10.1128/jvi.00144-23

    Figure Lengend Snippet: Antibodies used in the study

    Article Snippet: ACE2 goat , 1:1,000 , Bio-Techne , AF933.

    Techniques: