niv  (Millipore)


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  • 99
    Name:
    Nivalenol solution
    Description:
    Certan Vial
    Catalog Number:
    34131
    Price:
    None
    Applications:
    Nivalenol was used as standard to investigate in vitro mycotoxin binding of deoxynivalenol and nivalenol by adsorbent materials (activated carbon).
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    Structured Review

    Millipore niv
    Nivalenol solution
    Certan Vial
    https://www.bioz.com/result/niv/product/Millipore
    Average 99 stars, based on 13 article reviews
    Price from $9.99 to $1999.99
    niv - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Nipah Virus Infects Specific Subsets of Porcine Peripheral Blood Mononuclear Cells"

    Article Title: Nipah Virus Infects Specific Subsets of Porcine Peripheral Blood Mononuclear Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0030855

    Stimulation of T cells with PMA/ionomycin and/or NiV infection. Figs. 5.A–5.D illustrate changes in cell appearance of the CD6+ T cells after 18 hrs post infection with NiV at 0.1 moi. Formation of cell clusters in NiV infected cells suggests that the infection stimulates T lymphocytes. Appearance of non-stimulated T cells ( Fig. 5.A ) in comparison to NiV infected T cells ( Fig. 5.B ) and the PMA/ionomycin stimulated cells ( Fig. 5.C ). NiV infected PMA/ionomycin activated T cells ( Fig. 5.D ). Fig. 5.E Cytokine RNA profiles were determined 48 hrs post incubation/infection by quantitative RT-PCR. Gray columns represent ratio of PMA/ionomycin activated cells and non-stimulated T cells; white columns represent ratio of NiV infected cells compare to non-stimulated cells; black columns represent ratio of infected PMA/ionomycin activated cells compared to PMA/ionomycin activated T cells.
    Figure Legend Snippet: Stimulation of T cells with PMA/ionomycin and/or NiV infection. Figs. 5.A–5.D illustrate changes in cell appearance of the CD6+ T cells after 18 hrs post infection with NiV at 0.1 moi. Formation of cell clusters in NiV infected cells suggests that the infection stimulates T lymphocytes. Appearance of non-stimulated T cells ( Fig. 5.A ) in comparison to NiV infected T cells ( Fig. 5.B ) and the PMA/ionomycin stimulated cells ( Fig. 5.C ). NiV infected PMA/ionomycin activated T cells ( Fig. 5.D ). Fig. 5.E Cytokine RNA profiles were determined 48 hrs post incubation/infection by quantitative RT-PCR. Gray columns represent ratio of PMA/ionomycin activated cells and non-stimulated T cells; white columns represent ratio of NiV infected cells compare to non-stimulated cells; black columns represent ratio of infected PMA/ionomycin activated cells compared to PMA/ionomycin activated T cells.

    Techniques Used: Infection, Incubation, Quantitative RT-PCR

    2) Product Images from "A Cross-Reactive Humanized Monoclonal Antibody Targeting Fusion Glycoprotein Function Protects Ferrets Against Lethal Nipah Virus and Hendra Virus Infection"

    Article Title: A Cross-Reactive Humanized Monoclonal Antibody Targeting Fusion Glycoprotein Function Protects Ferrets Against Lethal Nipah Virus and Hendra Virus Infection

    Journal: The Journal of Infectious Diseases

    doi: 10.1093/infdis/jiz515

    (A) Kaplan-Meier survival curve for Nipah virus (NiV)-challenged ferrets treated with h5B3.1. Red NiV control (n = 1), green D1/D3 cohort (n = 3), and blue D3/D5 cohort (n = 3). Asterisk = NiV challenge day; green nabla, 5B3.1 treatment day for D1/D3 cohort; and blue nabla, treatment day for D3/D5 cohort. (B) Percentage weight from day 0 post-NiV challenge. Red NiV control (n = 1), green D1/D3 cohort (n = 3), and blue D3/D5 cohort (n = 3). (C) Plaque-forming units (PFU) per mL isolated from whole blood on indicated days after NiV challenge. (D) The PFU per gram of tissue at the study end point for each NiV-challenged ferret in the study. NiV-C = control; NiV-1–3 green, D1/D3 cohort; and NiV-4–6 blue, D3/5 cohort.
    Figure Legend Snippet: (A) Kaplan-Meier survival curve for Nipah virus (NiV)-challenged ferrets treated with h5B3.1. Red NiV control (n = 1), green D1/D3 cohort (n = 3), and blue D3/D5 cohort (n = 3). Asterisk = NiV challenge day; green nabla, 5B3.1 treatment day for D1/D3 cohort; and blue nabla, treatment day for D3/D5 cohort. (B) Percentage weight from day 0 post-NiV challenge. Red NiV control (n = 1), green D1/D3 cohort (n = 3), and blue D3/D5 cohort (n = 3). (C) Plaque-forming units (PFU) per mL isolated from whole blood on indicated days after NiV challenge. (D) The PFU per gram of tissue at the study end point for each NiV-challenged ferret in the study. NiV-C = control; NiV-1–3 green, D1/D3 cohort; and NiV-4–6 blue, D3/5 cohort.

    Techniques Used: Isolation

    Detection of specific anti-Nipah virus (NiV) G (A) and anti-HeV G (B) immunoglobulin G (IgG) antibodies circulating in ferrets at indicated days postvirus challenge. NiV-C = control; NiV-1–3 green, D1/D3 cohort; and NiV-4–6 blue, D3/5 cohort. HeV-C = control; and HeV-1–3 gray, D3/5 cohort. Mean fluorescence intensities (MFI) are shown on the y-axis and represent binding of specific IgG. Error bars represent the standard deviation of fluorescence intensity across 100 beads for each sample.
    Figure Legend Snippet: Detection of specific anti-Nipah virus (NiV) G (A) and anti-HeV G (B) immunoglobulin G (IgG) antibodies circulating in ferrets at indicated days postvirus challenge. NiV-C = control; NiV-1–3 green, D1/D3 cohort; and NiV-4–6 blue, D3/5 cohort. HeV-C = control; and HeV-1–3 gray, D3/5 cohort. Mean fluorescence intensities (MFI) are shown on the y-axis and represent binding of specific IgG. Error bars represent the standard deviation of fluorescence intensity across 100 beads for each sample.

    Techniques Used: Fluorescence, Binding Assay, Standard Deviation

    3) Product Images from "Simultaneous determination of major type A and B trichothecenes, zearalenone and certain modified metabolites in Finnish cereal grains with a novel liquid chromatography-tandem mass spectrometric method"

    Article Title: Simultaneous determination of major type A and B trichothecenes, zearalenone and certain modified metabolites in Finnish cereal grains with a novel liquid chromatography-tandem mass spectrometric method

    Journal: Analytical and Bioanalytical Chemistry

    doi: 10.1007/s00216-015-8676-4

    Overlayed quantifier extracted ion chromatograms of a blank wheat sample spiked at the medium level, with 1 NIV3Glc; 2 NIV; 3 DON3Glc; 4 DON; 5 β-ZEL14Glc; 6 ZEN16Glc; 7 α-ZEL14Glc; 8 HT2-3-Glc; 9 3Ac-DON; 10 ZEN14Glc; 11 ZEN14Sulf; 12 HT2; 13 β-ZEL; 14 α-ZEL; 15 T2; 16 ZEN
    Figure Legend Snippet: Overlayed quantifier extracted ion chromatograms of a blank wheat sample spiked at the medium level, with 1 NIV3Glc; 2 NIV; 3 DON3Glc; 4 DON; 5 β-ZEL14Glc; 6 ZEN16Glc; 7 α-ZEL14Glc; 8 HT2-3-Glc; 9 3Ac-DON; 10 ZEN14Glc; 11 ZEN14Sulf; 12 HT2; 13 β-ZEL; 14 α-ZEL; 15 T2; 16 ZEN

    Techniques Used: Gas Chromatography

    4) Product Images from "Attachment Protein G of an African Bat Henipavirus Is Differentially Restricted in Chiropteran and Nonchiropteran Cells"

    Article Title: Attachment Protein G of an African Bat Henipavirus Is Differentially Restricted in Chiropteran and Nonchiropteran Cells

    Journal: Journal of Virology

    doi: 10.1128/JVI.01561-14

    Expression of NiV-G and M74-G in permeabilized cells. (A) Vero76, EidNi/41, HypLu/2, and HypNi/1.1 cells were transfected for expression of FLAG-tagged NiV-G or M74-G. At 24 h p.t., permeabilized cells were immunostained for the presence of FLAG-tagged
    Figure Legend Snippet: Expression of NiV-G and M74-G in permeabilized cells. (A) Vero76, EidNi/41, HypLu/2, and HypNi/1.1 cells were transfected for expression of FLAG-tagged NiV-G or M74-G. At 24 h p.t., permeabilized cells were immunostained for the presence of FLAG-tagged

    Techniques Used: Expressing, Transfection

    Surface expression of M74-G. (A) Cells were transfected for expression of FLAG-tagged NiV-G or M74-G. At 24 h p.t., permeabilized (+TX) and nonpermeabilized (−TX) cells were immunostained by antibodies directed against the FLAG tag. Scale bars
    Figure Legend Snippet: Surface expression of M74-G. (A) Cells were transfected for expression of FLAG-tagged NiV-G or M74-G. At 24 h p.t., permeabilized (+TX) and nonpermeabilized (−TX) cells were immunostained by antibodies directed against the FLAG tag. Scale bars

    Techniques Used: Expressing, Transfection, FLAG-tag

    Expression of chimeric G proteins of NiV and M74. (A) HypNi/1.1 cells were transfected for coexpression of NiV-G-HA (green) with either M74-G ED or NiV-G ED (red). Permeabilized cells were immunostained for NiV-G with antibodies against the HA tag (green)
    Figure Legend Snippet: Expression of chimeric G proteins of NiV and M74. (A) HypNi/1.1 cells were transfected for coexpression of NiV-G-HA (green) with either M74-G ED or NiV-G ED (red). Permeabilized cells were immunostained for NiV-G with antibodies against the HA tag (green)

    Techniques Used: Expressing, Transfection

    Coexpression of F and G of NiV (A) and M74 (B) in HypLu/2, EidNi/41, and HBE cells. At 24 h p.t., cells were fixed, permeabilized, and incubated with anti-HA and anti-FLAG antibodies, followed by incubation with secondary Cy3- and FITC-conjugated antibodies.
    Figure Legend Snippet: Coexpression of F and G of NiV (A) and M74 (B) in HypLu/2, EidNi/41, and HBE cells. At 24 h p.t., cells were fixed, permeabilized, and incubated with anti-HA and anti-FLAG antibodies, followed by incubation with secondary Cy3- and FITC-conjugated antibodies.

    Techniques Used: Incubation

    Flow cytometry analysis of the surface expression of M74-G. Transfected cells expressing no foreign protein (pCG1) or FLAG-tagged NiV-G or M74-G were incubated with an antibody against the FLAG epitope, followed by incubation with anti-mouse biotin and
    Figure Legend Snippet: Flow cytometry analysis of the surface expression of M74-G. Transfected cells expressing no foreign protein (pCG1) or FLAG-tagged NiV-G or M74-G were incubated with an antibody against the FLAG epitope, followed by incubation with anti-mouse biotin and

    Techniques Used: Flow Cytometry, Cytometry, Expressing, Transfection, Incubation, FLAG-tag

    5) Product Images from "Nipah Virus Uses Leukocytes for Efficient Dissemination within a Host ▿"

    Article Title: Nipah Virus Uses Leukocytes for Efficient Dissemination within a Host ▿

    Journal: Journal of Virology

    doi: 10.1128/JVI.00549-11

    Transinfection of NiV ex vivo and in vivo . (A and B) Hamster splenocytes are not permissive to infection in vitro with rNiV-EGFP, as observed under light (A) and fluorescence (B) microscopes at 4 days p.i. (C to I) Hamsters were infected with 10 3 PFU
    Figure Legend Snippet: Transinfection of NiV ex vivo and in vivo . (A and B) Hamster splenocytes are not permissive to infection in vitro with rNiV-EGFP, as observed under light (A) and fluorescence (B) microscopes at 4 days p.i. (C to I) Hamsters were infected with 10 3 PFU

    Techniques Used: Ex Vivo, In Vivo, Infection, In Vitro, Fluorescence

    NiV replication in human leukocytes. (A) Kinetics of the expression of NiV N, M, F, G, and L genes in PBLs, DC, macrophages, and U373 cells infected with NiV after analysis by RT-qPCR. (B) NiV-EGFP replication in human cells 48 h p.i. observed under light
    Figure Legend Snippet: NiV replication in human leukocytes. (A) Kinetics of the expression of NiV N, M, F, G, and L genes in PBLs, DC, macrophages, and U373 cells infected with NiV after analysis by RT-qPCR. (B) NiV-EGFP replication in human cells 48 h p.i. observed under light

    Techniques Used: Expressing, Infection, Quantitative RT-PCR

    Transinfection capacity of human leukocytes. (A) Experimental protocol. PBLs, DC, or monocytes were infected with NiV-EGFP at either 37°C or 4°C in the presence or absence of EIPA, an inhibitor of NiV entry; washed; and returned to culture.
    Figure Legend Snippet: Transinfection capacity of human leukocytes. (A) Experimental protocol. PBLs, DC, or monocytes were infected with NiV-EGFP at either 37°C or 4°C in the presence or absence of EIPA, an inhibitor of NiV entry; washed; and returned to culture.

    Techniques Used: Infection

    6) Product Images from "Population Analysis of the Fusarium graminearum Species Complex from Wheat in China Show a Shift to More Aggressive Isolates"

    Article Title: Population Analysis of the Fusarium graminearum Species Complex from Wheat in China Show a Shift to More Aggressive Isolates

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0031722

    Genetic clusters compositions of F. graminearum s. str. with 15ADON chemotype (N = 169), F. asiaticum with 3ADON chemotype (N = 171) and with NIV chemotype (N = 97).
    Figure Legend Snippet: Genetic clusters compositions of F. graminearum s. str. with 15ADON chemotype (N = 169), F. asiaticum with 3ADON chemotype (N = 171) and with NIV chemotype (N = 97).

    Techniques Used:

    Visual representation of isolate distribution (A) and admixture estimates (B) based on VNTR data. A: green = F. graminearum s. str. with 15ADON type, red = F. asiaticum with 3ADON type, blue = F. asiaticum with NIV type, yellow = F. asiaticum with 15ADON type; B. Visual representation of admixture estimates based on VNTR data for 275 F. asiaticum and 169 F. graminearum s. str. isolates collected from three Chinese populations (3ADON, NIV and 15ADON). Each population is represented by a unique color (3ADON = red, NIV = blue and 15ADON = green). Individual isolates are represented by a distinct vertical line colored to represent the estimated proportion of the isolates genome derived from each population. The horizontal axis consists of a single vertical bar for each of 444 isolates. Isolates were assigned to a specific population when membership fraction ≥0.8.
    Figure Legend Snippet: Visual representation of isolate distribution (A) and admixture estimates (B) based on VNTR data. A: green = F. graminearum s. str. with 15ADON type, red = F. asiaticum with 3ADON type, blue = F. asiaticum with NIV type, yellow = F. asiaticum with 15ADON type; B. Visual representation of admixture estimates based on VNTR data for 275 F. asiaticum and 169 F. graminearum s. str. isolates collected from three Chinese populations (3ADON, NIV and 15ADON). Each population is represented by a unique color (3ADON = red, NIV = blue and 15ADON = green). Individual isolates are represented by a distinct vertical line colored to represent the estimated proportion of the isolates genome derived from each population. The horizontal axis consists of a single vertical bar for each of 444 isolates. Isolates were assigned to a specific population when membership fraction ≥0.8.

    Techniques Used: Derivative Assay

    Related Articles

    Injection:

    Article Title: Toxigenic potential of Fusarium graminearum isolated from maize of northwest Argentina
    Article Snippet: .. Trichothecenes were quantified using external standards of deoxynivalenol, nivalenol and acetylated forms of deoxynivalenol (Sigma-Aldrich Co. St Louis, MO) injected at concentrations of 1 to 4 μg mL−1 in acetonitrile/water (84:16). ..

    Incubation:

    Article Title: Attachment Protein G of an African Bat Henipavirus Is Differentially Restricted in Chiropteran and Nonchiropteran Cells
    Article Snippet: .. NiV- and M74-G-FLAG were detected by incubation with an anti-FLAG antibody (mouse; Sigma) and anti-mouse horseradish peroxidase (HRP; Dako). .. For the visualization of protein bands, membranes containing the immobilized proteins were incubated with Super Signal West Dura extended duration substrate (Thermo Scientific), placed in a ChemiDoc imager (Bio-Rad), and analyzed with the Quanti One software (Bio-Rad).

    Infection:

    Article Title: Nipah Virus Uses Leukocytes for Efficient Dissemination within a Host ▿
    Article Snippet: .. DC, PBLs, and monocytes were prepared from fresh blood and infected for 1 h with NiV or rNiV-EGFP at either 4°C or 37°C in the presence or absence of 50 μM 5-( N -ethyl- N -isopropyl) amiloride (EIPA) (Sigma). .. In some experiments, 106 PBLs were preincubated with 100 μg/ml of EphB4 soluble Fc fusion protein (ligand for ephrinB2; R & D Systems) for 45 min, with 1 mg/ml of pronase (Roche) or 2 mg/ml of trypsin (Sigma) for 30 min at 37°C, or with 5 mM EDTA or 5 mM EGTA (Sigma) for 10 min at room temperature; washed; and infected with NiV.

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  • 90
    Millipore mutant niv f
    Both the <t>NiV</t> G head and stalk regions alone are capable of interacting with NiV F. (A) Flow-cytometric plots of the NiV G constructs. PC, pcDNA3 vector-only control. G detection is shown on the y axis, and F detection is shown on the x axis. (B) Normalized G MFI values of the NiV G constructs determined by flow cytometry. Values are normalized to G ecto . Averages and standard deviations are shown ( n = 5). (C) Cell lysates of cells transfected with wt F and either wt or mutant G constructs. NiV G was blotted with rabbit anti-HA, and <t>NiV</t> F was blotted with mouse anti-AU1. Asterisks denote the G protein band. (D) Pulldown of NiV G and co-IP of F for transfected cells. Antibody blotting was performed as described for panel C. (E) Syncytium quantification of wt F transfected with wt or soluble G construct DNA. Values are normalized to those of wt G ( n = 3).
    Mutant Niv F, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mutant niv f/product/Millipore
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    85
    Millipore niv hr2 fc expression plasmid
    <t>NiV-F</t> CT fusion mutants are differentially resistant to fusion inhibition by NiV-F <t>HR2-Fc</t> and exhibit corresponding rates of fusion kinetics relative to WT NiV-F. (A) The sensitivity of NiV envelope-mediated fusion to inhibition by NiV-HR2-Fc is shown
    Niv Hr2 Fc Expression Plasmid, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore soluble niv f
    Architecture of the prefusion <t>NiV</t> F glycoprotein. a , b , Ribbon diagrams of a NiV F ectodomain protomer from the cryo-EM structure of NiV F in complex with the 5B3 Fab fragment. HRA, heptad-repeat A; HRB, heptad-repeat B.
    Soluble Niv F, supplied by Millipore, 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/soluble niv f/product/Millipore
    Average 93 stars, based on 1 article reviews
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    soluble niv f - by Bioz Stars, 2020-09
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    88
    Millipore niv b infected b
    Monocyte chemotaxis in NiV-M- and <t>NiV-B-infected</t> B- and S-ALI. Monocyte chemotaxis assay was performed using mixtures similar to the basal side of B- and S-ALI cultures (a), or samples made of only one cytokine or chemokine at a time (b), as described
    Niv B Infected B, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Both the NiV G head and stalk regions alone are capable of interacting with NiV F. (A) Flow-cytometric plots of the NiV G constructs. PC, pcDNA3 vector-only control. G detection is shown on the y axis, and F detection is shown on the x axis. (B) Normalized G MFI values of the NiV G constructs determined by flow cytometry. Values are normalized to G ecto . Averages and standard deviations are shown ( n = 5). (C) Cell lysates of cells transfected with wt F and either wt or mutant G constructs. NiV G was blotted with rabbit anti-HA, and NiV F was blotted with mouse anti-AU1. Asterisks denote the G protein band. (D) Pulldown of NiV G and co-IP of F for transfected cells. Antibody blotting was performed as described for panel C. (E) Syncytium quantification of wt F transfected with wt or soluble G construct DNA. Values are normalized to those of wt G ( n = 3).

    Journal: Journal of Virology

    Article Title: Multiple Strategies Reveal a Bidentate Interaction between the Nipah Virus Attachment and Fusion Glycoproteins

    doi: 10.1128/JVI.01469-16

    Figure Lengend Snippet: Both the NiV G head and stalk regions alone are capable of interacting with NiV F. (A) Flow-cytometric plots of the NiV G constructs. PC, pcDNA3 vector-only control. G detection is shown on the y axis, and F detection is shown on the x axis. (B) Normalized G MFI values of the NiV G constructs determined by flow cytometry. Values are normalized to G ecto . Averages and standard deviations are shown ( n = 5). (C) Cell lysates of cells transfected with wt F and either wt or mutant G constructs. NiV G was blotted with rabbit anti-HA, and NiV F was blotted with mouse anti-AU1. Asterisks denote the G protein band. (D) Pulldown of NiV G and co-IP of F for transfected cells. Antibody blotting was performed as described for panel C. (E) Syncytium quantification of wt F transfected with wt or soluble G construct DNA. Values are normalized to those of wt G ( n = 3).

    Article Snippet: Transfected 293T cells expressing wt or mutant NiV G in the presence or absence of wt NiV F or expressing wt NiV G in the presence or absence of wt or mutant NiV F were lysed in radioimmunoprecipitation assay (RIPA) buffer (Millipore) supplemented with complete protease inhibitor (cOmplete Mini; Roche).

    Techniques: Flow Cytometry, Construct, Plasmid Preparation, Cytometry, Transfection, Mutagenesis, Co-Immunoprecipitation Assay

    Overview of the NiV G and NiV F glycoproteins. (A) Schematic of NiV G. CT, cytoplasmic tail; TM, transmembrane domain; HA, hemagglutinin tag. Numbers represent amino acid residues. (B) Schematic of NiV F. HR, heptad repeat region; FP, fusion peptide. The dashed line represents the cleavage site. A cytoplasmic AU1 or F 2 subunit FLAG tag was used for detection. (C) Schematic of the soluble G ectodomain construct, G ecto . Igκ, immunoglobulin kappa light chain sequence. (D) Schematic of the soluble G head construct, G head . (E) Schematic of the soluble G stalk construct, G stalk . (F) Schematic of the soluble F ectodomain construct F tri . The GCNt motif represents a trimeric coiled-coil domain.

    Journal: Journal of Virology

    Article Title: Multiple Strategies Reveal a Bidentate Interaction between the Nipah Virus Attachment and Fusion Glycoproteins

    doi: 10.1128/JVI.01469-16

    Figure Lengend Snippet: Overview of the NiV G and NiV F glycoproteins. (A) Schematic of NiV G. CT, cytoplasmic tail; TM, transmembrane domain; HA, hemagglutinin tag. Numbers represent amino acid residues. (B) Schematic of NiV F. HR, heptad repeat region; FP, fusion peptide. The dashed line represents the cleavage site. A cytoplasmic AU1 or F 2 subunit FLAG tag was used for detection. (C) Schematic of the soluble G ectodomain construct, G ecto . Igκ, immunoglobulin kappa light chain sequence. (D) Schematic of the soluble G head construct, G head . (E) Schematic of the soluble G stalk construct, G stalk . (F) Schematic of the soluble F ectodomain construct F tri . The GCNt motif represents a trimeric coiled-coil domain.

    Article Snippet: Transfected 293T cells expressing wt or mutant NiV G in the presence or absence of wt NiV F or expressing wt NiV G in the presence or absence of wt or mutant NiV F were lysed in radioimmunoprecipitation assay (RIPA) buffer (Millipore) supplemented with complete protease inhibitor (cOmplete Mini; Roche).

    Techniques: FLAG-tag, Construct, Sequencing

    Models for G-F interactions leading to F triggering. (A) Prior to ephrinB2 (blue) receptor binding, the head region of NiV G (purple) interacts (represented by red lines) with NiV F (green) while the C-terminal domain of the stalk is relatively covered. (B) Upon receptor binding, the G head undergoes conformational changes that result in exposure of the C-terminal portion of the stalk (yellow), resulting in the G-F interactions switching from the G head to the G stalk. (C) Interaction of the G stalk C-terminal domain with F triggers F to undergo its own conformational changes, which execute virus-host cell membrane fusion. (D) Alternatively, both the head and N-terminal stalk regions interact with F prior to receptor binding, after which conformational changes in G result in exposure of the G stalk C-terminal domain, which triggers F and executes membrane fusion as shown in panels B and C. For simplicity, ephrinB2, NiV G, and NiV F are represented as monomers.

    Journal: Journal of Virology

    Article Title: Multiple Strategies Reveal a Bidentate Interaction between the Nipah Virus Attachment and Fusion Glycoproteins

    doi: 10.1128/JVI.01469-16

    Figure Lengend Snippet: Models for G-F interactions leading to F triggering. (A) Prior to ephrinB2 (blue) receptor binding, the head region of NiV G (purple) interacts (represented by red lines) with NiV F (green) while the C-terminal domain of the stalk is relatively covered. (B) Upon receptor binding, the G head undergoes conformational changes that result in exposure of the C-terminal portion of the stalk (yellow), resulting in the G-F interactions switching from the G head to the G stalk. (C) Interaction of the G stalk C-terminal domain with F triggers F to undergo its own conformational changes, which execute virus-host cell membrane fusion. (D) Alternatively, both the head and N-terminal stalk regions interact with F prior to receptor binding, after which conformational changes in G result in exposure of the G stalk C-terminal domain, which triggers F and executes membrane fusion as shown in panels B and C. For simplicity, ephrinB2, NiV G, and NiV F are represented as monomers.

    Article Snippet: Transfected 293T cells expressing wt or mutant NiV G in the presence or absence of wt NiV F or expressing wt NiV G in the presence or absence of wt or mutant NiV F were lysed in radioimmunoprecipitation assay (RIPA) buffer (Millipore) supplemented with complete protease inhibitor (cOmplete Mini; Roche).

    Techniques: Binding Assay

    NiV-F CT fusion mutants are differentially resistant to fusion inhibition by NiV-F HR2-Fc and exhibit corresponding rates of fusion kinetics relative to WT NiV-F. (A) The sensitivity of NiV envelope-mediated fusion to inhibition by NiV-HR2-Fc is shown

    Journal:

    Article Title: Polybasic KKR Motif in the Cytoplasmic Tail of Nipah Virus Fusion Protein Modulates Membrane Fusion by Inside-Out Signaling ▿

    doi: 10.1128/JVI.02205-06

    Figure Lengend Snippet: NiV-F CT fusion mutants are differentially resistant to fusion inhibition by NiV-F HR2-Fc and exhibit corresponding rates of fusion kinetics relative to WT NiV-F. (A) The sensitivity of NiV envelope-mediated fusion to inhibition by NiV-HR2-Fc is shown

    Article Snippet: The NiV-HR2-Fc expression plasmid was transfected into 293T cells, and at 24 h posttransfection, supernatants were collected and concentrated with a Centriplus YM-10 filter (Millipore, Bedford, MA).

    Techniques: Inhibition

    Architecture of the prefusion NiV F glycoprotein. a , b , Ribbon diagrams of a NiV F ectodomain protomer from the cryo-EM structure of NiV F in complex with the 5B3 Fab fragment. HRA, heptad-repeat A; HRB, heptad-repeat B.

    Journal: Nature Structural & Molecular Biology

    Article Title: An antibody against the F glycoprotein inhibits Nipah and Hendra virus infections

    doi: 10.1038/s41594-019-0308-9

    Figure Lengend Snippet: Architecture of the prefusion NiV F glycoprotein. a , b , Ribbon diagrams of a NiV F ectodomain protomer from the cryo-EM structure of NiV F in complex with the 5B3 Fab fragment. HRA, heptad-repeat A; HRB, heptad-repeat B.

    Article Snippet: Soluble NiV F and HeV F were produced by transient transfection of FreeStyle 293F cells at a density of 1 × 106 cells ml−1 with the corresponding plasmid using 293-Free transfection reagent (Millipore) and Opti-MEM (Thermo-Fisher) according to the manufacturer’s protocol.

    Techniques:

    Cryo-EM characterization of the NiV F glycoprotein in complex with the neutralizing antibody 5B3 Fab fragment. a , Representative micrograph. Scale bar, 100 nm. b , Reference-free 2D class averages. Scale bar, 100 Å. c , Gold-standard (black) and map/model (red) Fourier shell correlation curves. Dotted lines indicate 0.143 and 0.5 thresholds. d , Two orthogonal views of the cryo-EM reconstruction colored by local resolution computed using cryoSPARC. e , Enlarged view of the model with the cryo-EM reconstruction rendered as a blue mesh.

    Journal: Nature Structural & Molecular Biology

    Article Title: An antibody against the F glycoprotein inhibits Nipah and Hendra virus infections

    doi: 10.1038/s41594-019-0308-9

    Figure Lengend Snippet: Cryo-EM characterization of the NiV F glycoprotein in complex with the neutralizing antibody 5B3 Fab fragment. a , Representative micrograph. Scale bar, 100 nm. b , Reference-free 2D class averages. Scale bar, 100 Å. c , Gold-standard (black) and map/model (red) Fourier shell correlation curves. Dotted lines indicate 0.143 and 0.5 thresholds. d , Two orthogonal views of the cryo-EM reconstruction colored by local resolution computed using cryoSPARC. e , Enlarged view of the model with the cryo-EM reconstruction rendered as a blue mesh.

    Article Snippet: Soluble NiV F and HeV F were produced by transient transfection of FreeStyle 293F cells at a density of 1 × 106 cells ml−1 with the corresponding plasmid using 293-Free transfection reagent (Millipore) and Opti-MEM (Thermo-Fisher) according to the manufacturer’s protocol.

    Techniques:

    The 5B3 and h5B3.1 neutralizing antibodies inhibit fusion by locking NiV F in the prefusion state. a , Molecular surface representation of the NiV F prefusion trimer showing the 5B3 footprint colored violet. b (PDB 1ZTM). c , 5B3 concentration-dependent inhibition of streptavidin-mediated pulldown of a biotinylated HRB/NiV F conformational intermediate complex in a triggering assay. The non-neutralizing control antibody (13G5), which is specific for postfusion F, had no effect. d , NiV F triggering assay carried out in the presence of 5B3 or h5B3.1 IgGs showing both mAbs prevented F fusogenic conformational changes. Subsequent protein G immunoprecipitation of F/5B3 and F/h5B3.1 complexes that were not pulled down by the biotinylated HRB peptide indicated the antibodies remain bound to F. e , Capture of an F fusogenic conformational intermediate could be partially rescued by raising the temperature to ≥60 °C, as detected by comparing streptavidin-mediated pulldown and protein G immunoprecipitation of F/5B3 and F/h5B3.1 complexes. In a and b , a single 5B3 epitope is colored for clarity. In all panels, precipitated samples were analyzed by western blotting using a rabbit anti-F antibody. f , g , NiV F ( f ) or HeV F ( g ) mediated cell-cell fusion could be inhibited by 5B3 or h5B3.1 in a concentration-dependent manner. D54 is an HIV envelope antibody used as negative control. Data shown are mean and s.d. for n = 2 technical replicates. Uncropped images for d and e online.

    Journal: Nature Structural & Molecular Biology

    Article Title: An antibody against the F glycoprotein inhibits Nipah and Hendra virus infections

    doi: 10.1038/s41594-019-0308-9

    Figure Lengend Snippet: The 5B3 and h5B3.1 neutralizing antibodies inhibit fusion by locking NiV F in the prefusion state. a , Molecular surface representation of the NiV F prefusion trimer showing the 5B3 footprint colored violet. b (PDB 1ZTM). c , 5B3 concentration-dependent inhibition of streptavidin-mediated pulldown of a biotinylated HRB/NiV F conformational intermediate complex in a triggering assay. The non-neutralizing control antibody (13G5), which is specific for postfusion F, had no effect. d , NiV F triggering assay carried out in the presence of 5B3 or h5B3.1 IgGs showing both mAbs prevented F fusogenic conformational changes. Subsequent protein G immunoprecipitation of F/5B3 and F/h5B3.1 complexes that were not pulled down by the biotinylated HRB peptide indicated the antibodies remain bound to F. e , Capture of an F fusogenic conformational intermediate could be partially rescued by raising the temperature to ≥60 °C, as detected by comparing streptavidin-mediated pulldown and protein G immunoprecipitation of F/5B3 and F/h5B3.1 complexes. In a and b , a single 5B3 epitope is colored for clarity. In all panels, precipitated samples were analyzed by western blotting using a rabbit anti-F antibody. f , g , NiV F ( f ) or HeV F ( g ) mediated cell-cell fusion could be inhibited by 5B3 or h5B3.1 in a concentration-dependent manner. D54 is an HIV envelope antibody used as negative control. Data shown are mean and s.d. for n = 2 technical replicates. Uncropped images for d and e online.

    Article Snippet: Soluble NiV F and HeV F were produced by transient transfection of FreeStyle 293F cells at a density of 1 × 106 cells ml−1 with the corresponding plasmid using 293-Free transfection reagent (Millipore) and Opti-MEM (Thermo-Fisher) according to the manufacturer’s protocol.

    Techniques: Concentration Assay, Inhibition, Immunoprecipitation, Western Blot, Negative Control

    Validation of the binding epitope on NiV F. a , Analysis of h5B3 scFv binding to full-length NiV F. scFv chimeric constructs in which the variable heavy h5B3 chain (VH 5B3 ), the variable light h5B3 chain (VL 5B3 ) or both chains were replaced with unrelated chains (VH h VL h ) from a human scFv library were assessed for binding to secreted wild-type NiV F. VH 5B3 /VL 5B3 scFv was used as a positive control. Western blotting was carried out using an anti-F polyclonal antibody to detect NiV F or anti-S-peptide antibody to detect the scFv. b , Site-directed mutagenesis of the 5B3 epitope. A panel of S-peptide tagged NiV F ectdomain mutants were generated and expressed in HEK 293T cells. The F-expressing cell lysates were divided equally and incubated with 5B3, 12B2 or S-protein agarose before immunoprecipitation/pulldown. Samples in which mAb 5B3 or 12B2 were added were precipitated with protein G Sepharose. Western blotting detection of the precipitated products was carried out using an anti-S-peptide antibody. The Gly247Ala substitution was used as positive control. c , Cell–cell fusion mediated by the NiV F mutants shown in b . Data are the mean percentage of wild-type fusion levels for each mutant normalized relative to total F expression, as measured by densitometry of western blot bands. The bars represent the standard error from three separate experiments.

    Journal: Nature Structural & Molecular Biology

    Article Title: An antibody against the F glycoprotein inhibits Nipah and Hendra virus infections

    doi: 10.1038/s41594-019-0308-9

    Figure Lengend Snippet: Validation of the binding epitope on NiV F. a , Analysis of h5B3 scFv binding to full-length NiV F. scFv chimeric constructs in which the variable heavy h5B3 chain (VH 5B3 ), the variable light h5B3 chain (VL 5B3 ) or both chains were replaced with unrelated chains (VH h VL h ) from a human scFv library were assessed for binding to secreted wild-type NiV F. VH 5B3 /VL 5B3 scFv was used as a positive control. Western blotting was carried out using an anti-F polyclonal antibody to detect NiV F or anti-S-peptide antibody to detect the scFv. b , Site-directed mutagenesis of the 5B3 epitope. A panel of S-peptide tagged NiV F ectdomain mutants were generated and expressed in HEK 293T cells. The F-expressing cell lysates were divided equally and incubated with 5B3, 12B2 or S-protein agarose before immunoprecipitation/pulldown. Samples in which mAb 5B3 or 12B2 were added were precipitated with protein G Sepharose. Western blotting detection of the precipitated products was carried out using an anti-S-peptide antibody. The Gly247Ala substitution was used as positive control. c , Cell–cell fusion mediated by the NiV F mutants shown in b . Data are the mean percentage of wild-type fusion levels for each mutant normalized relative to total F expression, as measured by densitometry of western blot bands. The bars represent the standard error from three separate experiments.

    Article Snippet: Soluble NiV F and HeV F were produced by transient transfection of FreeStyle 293F cells at a density of 1 × 106 cells ml−1 with the corresponding plasmid using 293-Free transfection reagent (Millipore) and Opti-MEM (Thermo-Fisher) according to the manufacturer’s protocol.

    Techniques: Binding Assay, Construct, Positive Control, Western Blot, Mutagenesis, Generated, Expressing, Incubation, Immunoprecipitation

    The 5B3 and h5B3.1 antibodies cross-react with NiV F and HeV F and potently neutralize NiV and HeV. a – d , Binding of 5B3 ( a , b ) or h5B3.1 ( c , d ) Fab fragments to immobilized NiV F ( a , c ) or HeV F ( b , d ) analyzed by biolayer interferometry. The concentrations of injected 5B3 and h5B3.1 are indicated in b and d , respectively. Fitted curves are shown as black dashed lines. The vertical dotted lines correspond to the transition between the association and dissociation phases. The experiments were performed in replicates with two different preparations of NiV F and of HeV F and a representative experiment is shown. e – g , Neutralization of authentic NiV-Malaysia (NiV-M) ( e ), NiV-Bangladesh (NiV-B) ( f ) and HeV ( g ) by the 5B3 (grey) and h5B3.1 (brown) immunoglobulin-Gs (IgGs). D10 (green) is an anti-HIV antibody used as negative control. Data shown are mean and s.d. for n = 3 independent experiments (with independent virus preparations).

    Journal: Nature Structural & Molecular Biology

    Article Title: An antibody against the F glycoprotein inhibits Nipah and Hendra virus infections

    doi: 10.1038/s41594-019-0308-9

    Figure Lengend Snippet: The 5B3 and h5B3.1 antibodies cross-react with NiV F and HeV F and potently neutralize NiV and HeV. a – d , Binding of 5B3 ( a , b ) or h5B3.1 ( c , d ) Fab fragments to immobilized NiV F ( a , c ) or HeV F ( b , d ) analyzed by biolayer interferometry. The concentrations of injected 5B3 and h5B3.1 are indicated in b and d , respectively. Fitted curves are shown as black dashed lines. The vertical dotted lines correspond to the transition between the association and dissociation phases. The experiments were performed in replicates with two different preparations of NiV F and of HeV F and a representative experiment is shown. e – g , Neutralization of authentic NiV-Malaysia (NiV-M) ( e ), NiV-Bangladesh (NiV-B) ( f ) and HeV ( g ) by the 5B3 (grey) and h5B3.1 (brown) immunoglobulin-Gs (IgGs). D10 (green) is an anti-HIV antibody used as negative control. Data shown are mean and s.d. for n = 3 independent experiments (with independent virus preparations).

    Article Snippet: Soluble NiV F and HeV F were produced by transient transfection of FreeStyle 293F cells at a density of 1 × 106 cells ml−1 with the corresponding plasmid using 293-Free transfection reagent (Millipore) and Opti-MEM (Thermo-Fisher) according to the manufacturer’s protocol.

    Techniques: Binding Assay, Injection, Neutralization, Negative Control

    5B3 binding is associated with a local structural reorganization of the HRA β-hairpin. a , Ribbon diagrams of the superimposed 5B3-bound and apo NiV F trimers. The 5B3 Fab fragments are omitted for clarity. The cyan square highlights the region of the structure shown in b – e . b , c , Enlarged views showing the HRA conformational change. d , e , Enlarged views rotated 45° relative to b and c . In all panels, 5B3-bound and apo-NiV F trimers are rendered grey and orange, respectively. In c – e (PDB 5EVM).

    Journal: Nature Structural & Molecular Biology

    Article Title: An antibody against the F glycoprotein inhibits Nipah and Hendra virus infections

    doi: 10.1038/s41594-019-0308-9

    Figure Lengend Snippet: 5B3 binding is associated with a local structural reorganization of the HRA β-hairpin. a , Ribbon diagrams of the superimposed 5B3-bound and apo NiV F trimers. The 5B3 Fab fragments are omitted for clarity. The cyan square highlights the region of the structure shown in b – e . b , c , Enlarged views showing the HRA conformational change. d , e , Enlarged views rotated 45° relative to b and c . In all panels, 5B3-bound and apo-NiV F trimers are rendered grey and orange, respectively. In c – e (PDB 5EVM).

    Article Snippet: Soluble NiV F and HeV F were produced by transient transfection of FreeStyle 293F cells at a density of 1 × 106 cells ml−1 with the corresponding plasmid using 293-Free transfection reagent (Millipore) and Opti-MEM (Thermo-Fisher) according to the manufacturer’s protocol.

    Techniques: Binding Assay

    The 5B3 neutralizing antibody recognizes a conserved quaternary epitope on the NiV F glycoprotein. a , Ribbon diagram of the NiV F trimer in complex with the 5B3 Fab fragment. One F protomer is rendered in teal and the other two protomers in grey. Only one Fab fragment is shown for clarity. b , Molecular surface representation of the NiV F trimer with the 5B3 CDR loops shown as ribbons, highlighting the quaternary nature of the epitope. c , Enlarged view of the interface between NiV F and 5B3 with selected residues rendered as sticks. NiV F residues are colored teal with oxygen and nitrogen atoms colored red and blue, respectively. In a – c , the 5B3 variable heavy (VH 5B3 ) and light (VL 5B3 ) chains are colored purple and pink, respectively. d , Molecular surface representation of the NiV F trimer showing the 5B3 footprint colored by residue conservation among NiV F and HeV F glycoproteins. Conservative sub.: conservative substitution; semi-conserv. sub.: semi-conservative substitution.

    Journal: Nature Structural & Molecular Biology

    Article Title: An antibody against the F glycoprotein inhibits Nipah and Hendra virus infections

    doi: 10.1038/s41594-019-0308-9

    Figure Lengend Snippet: The 5B3 neutralizing antibody recognizes a conserved quaternary epitope on the NiV F glycoprotein. a , Ribbon diagram of the NiV F trimer in complex with the 5B3 Fab fragment. One F protomer is rendered in teal and the other two protomers in grey. Only one Fab fragment is shown for clarity. b , Molecular surface representation of the NiV F trimer with the 5B3 CDR loops shown as ribbons, highlighting the quaternary nature of the epitope. c , Enlarged view of the interface between NiV F and 5B3 with selected residues rendered as sticks. NiV F residues are colored teal with oxygen and nitrogen atoms colored red and blue, respectively. In a – c , the 5B3 variable heavy (VH 5B3 ) and light (VL 5B3 ) chains are colored purple and pink, respectively. d , Molecular surface representation of the NiV F trimer showing the 5B3 footprint colored by residue conservation among NiV F and HeV F glycoproteins. Conservative sub.: conservative substitution; semi-conserv. sub.: semi-conservative substitution.

    Article Snippet: Soluble NiV F and HeV F were produced by transient transfection of FreeStyle 293F cells at a density of 1 × 106 cells ml−1 with the corresponding plasmid using 293-Free transfection reagent (Millipore) and Opti-MEM (Thermo-Fisher) according to the manufacturer’s protocol.

    Techniques:

    Monocyte chemotaxis in NiV-M- and NiV-B-infected B- and S-ALI. Monocyte chemotaxis assay was performed using mixtures similar to the basal side of B- and S-ALI cultures (a), or samples made of only one cytokine or chemokine at a time (b), as described

    Journal: The Journal of General Virology

    Article Title: Characterization of Nipah virus infection in a model of human airway epithelial cells cultured at an air–liquid interface

    doi: 10.1099/jgv.0.000441

    Figure Lengend Snippet: Monocyte chemotaxis in NiV-M- and NiV-B-infected B- and S-ALI. Monocyte chemotaxis assay was performed using mixtures similar to the basal side of B- and S-ALI cultures (a), or samples made of only one cytokine or chemokine at a time (b), as described

    Article Snippet: Cytokine/chemokine concentrations in the supernatant of NiV-M- and NiV-B-infected B- and S-ALI were determined using a Milliplex Human Cytokine 15 Plex Immunoassay custom kit (Millipore) and a Verikine-HS Human IFN Beta ELISA kit (PBL Assay Science).

    Techniques: Chemotaxis Assay, Infection