mers s  (Sino Biological)


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    Name:
    MERS CoV Spike Protein S1 Antibody Rabbit PAb
    Description:
    Produced in rabbits immunized with purified a synthetic peptide corresponding to the center region of the S1 subunit of MERS CoV NCoV Novel coronavirus spike glycoprotein S protein and purified by antigen affinity chromatography
    Catalog Number:
    100207-RP02
    Price:
    None
    Category:
    Primary Antibody
    Reactivity:
    MERS CoV
    Applications:
    WB
    Immunogen:
    A synthetic peptide corresponding to the center region of the S1 subunit of MERS-CoV (NCoV / Novel coronavirus) spike glycoprotein (S protein).
    Product Aliases:
    Anti-coronavirus s1 Antibody, Anti-coronavirus s2 Antibody, Anti-coronavirus spike Antibody, Anti-cov spike Antibody, Anti-ncov RBD Antibody, Anti-ncov s1 Antibody, Anti-ncov s2 Antibody, Anti-ncov spike Antibody, Anti-RBD Antibody, Anti-S Antibody, Anti-s1 Antibody, Anti-Spike RBD Antibody
    Antibody Type:
    PAb
    Host:
    Rabbit
    Isotype:
    Rabbit IgG
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    Structured Review

    Sino Biological mers s
    RXXR motifs located at the border between S1 and <t>S2</t> are required for efficient processing of the Middle East respiratory syndrome coronavirus spike protein <t>(MERS-S).</t> A , The domain organization of the MERS-S protein is schematically depicted. The MERS-S sequence at the border between the S1 and S2 subunits is shown. RXXR motifs, which constitute potential cleavage sites, are highlighted, and the predicted start of the S2 subunit is underlined. The mutations introduced into the potential cleavage sites in MERS-S are shown. B , 293T cells were transfected with expression plasmids coding for MERS-S wild type and the indicated MERS-S mutants equipped with a C-terminal V5 tag. Transfection of empty plasmid (pcDNA) served as negative control. Expression of S proteins in cell lysates was determined by Western blot, using a V5 tag–specific monoclonal antibody. Expression of β-actin in cell lysates was assessed as a loading control. The results shown are representative for at least 3 independent experiments. Abbreviations: CT, cytoplasmic tail; PCM, potential cleavage site mutant; RBD, receptor binding domain; SP, signal peptide; TM, transmembrane domain.
    Produced in rabbits immunized with purified a synthetic peptide corresponding to the center region of the S1 subunit of MERS CoV NCoV Novel coronavirus spike glycoprotein S protein and purified by antigen affinity chromatography
    https://www.bioz.com/result/mers s/product/Sino Biological
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mers s - by Bioz Stars, 2021-06
    93/100 stars

    Images

    1) Product Images from "Inhibition of Proprotein Convertases Abrogates Processing of the Middle Eastern Respiratory Syndrome Coronavirus Spike Protein in Infected Cells but Does Not Reduce Viral Infectivity"

    Article Title: Inhibition of Proprotein Convertases Abrogates Processing of the Middle Eastern Respiratory Syndrome Coronavirus Spike Protein in Infected Cells but Does Not Reduce Viral Infectivity

    Journal: The Journal of Infectious Diseases

    doi: 10.1093/infdis/jiu407

    RXXR motifs located at the border between S1 and S2 are required for efficient processing of the Middle East respiratory syndrome coronavirus spike protein (MERS-S). A , The domain organization of the MERS-S protein is schematically depicted. The MERS-S sequence at the border between the S1 and S2 subunits is shown. RXXR motifs, which constitute potential cleavage sites, are highlighted, and the predicted start of the S2 subunit is underlined. The mutations introduced into the potential cleavage sites in MERS-S are shown. B , 293T cells were transfected with expression plasmids coding for MERS-S wild type and the indicated MERS-S mutants equipped with a C-terminal V5 tag. Transfection of empty plasmid (pcDNA) served as negative control. Expression of S proteins in cell lysates was determined by Western blot, using a V5 tag–specific monoclonal antibody. Expression of β-actin in cell lysates was assessed as a loading control. The results shown are representative for at least 3 independent experiments. Abbreviations: CT, cytoplasmic tail; PCM, potential cleavage site mutant; RBD, receptor binding domain; SP, signal peptide; TM, transmembrane domain.
    Figure Legend Snippet: RXXR motifs located at the border between S1 and S2 are required for efficient processing of the Middle East respiratory syndrome coronavirus spike protein (MERS-S). A , The domain organization of the MERS-S protein is schematically depicted. The MERS-S sequence at the border between the S1 and S2 subunits is shown. RXXR motifs, which constitute potential cleavage sites, are highlighted, and the predicted start of the S2 subunit is underlined. The mutations introduced into the potential cleavage sites in MERS-S are shown. B , 293T cells were transfected with expression plasmids coding for MERS-S wild type and the indicated MERS-S mutants equipped with a C-terminal V5 tag. Transfection of empty plasmid (pcDNA) served as negative control. Expression of S proteins in cell lysates was determined by Western blot, using a V5 tag–specific monoclonal antibody. Expression of β-actin in cell lysates was assessed as a loading control. The results shown are representative for at least 3 independent experiments. Abbreviations: CT, cytoplasmic tail; PCM, potential cleavage site mutant; RBD, receptor binding domain; SP, signal peptide; TM, transmembrane domain.

    Techniques Used: Sequencing, Transfection, Expressing, Plasmid Preparation, Negative Control, Western Blot, Mutagenesis, Binding Assay

    Proprotein convertase activity is dispensable for Middle East respiratory syndrome coronavirus spike protein (MERS-S)–driven cell-cell and virus-cell fusion. A , Fusion of 293T effector cells transfected to express MERS-S with target cells transfected to express DPP4 and/or TMPRSS2 or control transfected with empty plasmid (pcDNA) was assessed. Both effector and target cells were incubated with 1 µM of proprotein convertase inhibitor (PCI) as indicated and, 1 day later, were mixed for cocultivation. The effector/target cell mixtures were incubated with phosphate-buffered saline (PBS) or PCI, and cell-cell fusion was quantified by determination of luciferase activities in cell lysates. The results of a representative experiment performed with triplicate samples are shown. Error bars indicate standard deviation (SD). Two separate experiments yielded similar results. B , Lentiviral pseudotypes carrying MERS-S, Lassa virus glycoprotein (LASV-GPC), Zaire ebolavirus glycoprotein (EBOV-GP), or the glycoprotein of vesicular stomatitis virus (VSV-G), as well as pseudotypes bearing no viral glycoprotein (pcDNA), were generated in the presence or absence of PCI (1 µM). Subsequently, target 293T cells transfected with empty plasmid or DPP4 expression plasmid were preincubated with dimethyl sulfoxide (DMSO), PBS, or PCI at a final concentration of 0.5 µM for 30–60 minutes, followed by transduction with the pseudotypes specified above. At 72 hours after transduction, luciferase activities in cell lysates were measured. The average of 5–7 independent experiments performed with triplicate samples is shown. Transduction with pseudotypes bearing VSV-G in the absence of inhibitor was set as 100%. Error bars indicate standard error of the mean (SEM). A 2-tailed Student t test was used to assess statistical significance.
    Figure Legend Snippet: Proprotein convertase activity is dispensable for Middle East respiratory syndrome coronavirus spike protein (MERS-S)–driven cell-cell and virus-cell fusion. A , Fusion of 293T effector cells transfected to express MERS-S with target cells transfected to express DPP4 and/or TMPRSS2 or control transfected with empty plasmid (pcDNA) was assessed. Both effector and target cells were incubated with 1 µM of proprotein convertase inhibitor (PCI) as indicated and, 1 day later, were mixed for cocultivation. The effector/target cell mixtures were incubated with phosphate-buffered saline (PBS) or PCI, and cell-cell fusion was quantified by determination of luciferase activities in cell lysates. The results of a representative experiment performed with triplicate samples are shown. Error bars indicate standard deviation (SD). Two separate experiments yielded similar results. B , Lentiviral pseudotypes carrying MERS-S, Lassa virus glycoprotein (LASV-GPC), Zaire ebolavirus glycoprotein (EBOV-GP), or the glycoprotein of vesicular stomatitis virus (VSV-G), as well as pseudotypes bearing no viral glycoprotein (pcDNA), were generated in the presence or absence of PCI (1 µM). Subsequently, target 293T cells transfected with empty plasmid or DPP4 expression plasmid were preincubated with dimethyl sulfoxide (DMSO), PBS, or PCI at a final concentration of 0.5 µM for 30–60 minutes, followed by transduction with the pseudotypes specified above. At 72 hours after transduction, luciferase activities in cell lysates were measured. The average of 5–7 independent experiments performed with triplicate samples is shown. Transduction with pseudotypes bearing VSV-G in the absence of inhibitor was set as 100%. Error bars indicate standard error of the mean (SEM). A 2-tailed Student t test was used to assess statistical significance.

    Techniques Used: Activity Assay, Transfection, Plasmid Preparation, Incubation, Luciferase, Standard Deviation, Gel Permeation Chromatography, Generated, Expressing, Concentration Assay, Transduction

    The Middle East respiratory syndrome coronavirus spike protein (MERS-S) is cleaved by proprotein convertases. 293T cells were transfected with expression plasmids encoding MERS-S, Zaire ebolavirus glycoprotein (EBOV-GP), or Lassa virus glycoprotein (LASV-GPC), all equipped with a C-terminal V5 tag. Cells transfected with empty plasmid (pcDNA) served as negative control. Subsequently, cells were incubated with the indicated concentrations of the proprotein convertase inhibitor (PCI). At 48 hours after transfection, glycoprotein expression was analyzed by Western blot, using a V5 tag–specific monoclonal antibody. Detection of β-actin served as loading control. The results shown are representative of three independent experiments.
    Figure Legend Snippet: The Middle East respiratory syndrome coronavirus spike protein (MERS-S) is cleaved by proprotein convertases. 293T cells were transfected with expression plasmids encoding MERS-S, Zaire ebolavirus glycoprotein (EBOV-GP), or Lassa virus glycoprotein (LASV-GPC), all equipped with a C-terminal V5 tag. Cells transfected with empty plasmid (pcDNA) served as negative control. Subsequently, cells were incubated with the indicated concentrations of the proprotein convertase inhibitor (PCI). At 48 hours after transfection, glycoprotein expression was analyzed by Western blot, using a V5 tag–specific monoclonal antibody. Detection of β-actin served as loading control. The results shown are representative of three independent experiments.

    Techniques Used: Transfection, Expressing, Gel Permeation Chromatography, Plasmid Preparation, Negative Control, Incubation, Western Blot

    The Middle East respiratory syndrome coronavirus (MERS-CoV) spike protein (MERS-S) is cleaved in transfected and infected cells. 293T cells were transfected with a plasmid encoding the MERS-S protein or with empty plasmid (pcDNA). Vero B4 cells were either infected with MERS-CoV at a multiplicity of infection of 5 or mock infected. Subsequently, the cells were lysed and analyzed by Western blot, using a polyclonal antibody directed against the S2 subunit of MERS-S. A β-actin antibody served as a loading control. Similar results were obtained in 2 separate experiments.
    Figure Legend Snippet: The Middle East respiratory syndrome coronavirus (MERS-CoV) spike protein (MERS-S) is cleaved in transfected and infected cells. 293T cells were transfected with a plasmid encoding the MERS-S protein or with empty plasmid (pcDNA). Vero B4 cells were either infected with MERS-CoV at a multiplicity of infection of 5 or mock infected. Subsequently, the cells were lysed and analyzed by Western blot, using a polyclonal antibody directed against the S2 subunit of MERS-S. A β-actin antibody served as a loading control. Similar results were obtained in 2 separate experiments.

    Techniques Used: Transfection, Infection, Plasmid Preparation, Western Blot

    2) Product Images from "Single-Dose, Intranasal Immunization with Recombinant Parainfluenza Virus 5 Expressing Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Spike Protein Protects Mice from Fatal MERS-CoV Infection"

    Article Title: Single-Dose, Intranasal Immunization with Recombinant Parainfluenza Virus 5 Expressing Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Spike Protein Protects Mice from Fatal MERS-CoV Infection

    Journal: mBio

    doi: 10.1128/mBio.00554-20

    Single-dose intranasal immunization with PIV5-MERS-S induced robust MERS-CoV-specific CD8 T cell response in human DPP4 knockin (hDPP4 KI) mice. (A) Schematic diagram showing the experimental plan to examine CD8 T cell response after immunization and challenge. hDPP4 KI mice were intranasally immunized with 10 4 PFU PIV5-MERS-S or PIV5-GFP. At 4 weeks or 4 days following immunization, mice were challenged with MERS MA 6.1.2, single-cell suspensions from the lungs of immunized mice were stimulated with MERS-CoV spike peptides (S343 and S1165), and specific CD8 T cells were determined by IFN-γ intracellular staining. (B to D) Representative FACS plots (B), percentage (C), and total number (D) of MERS-CoV-specific CD8 T cells in the lungs at 4 weeks after immunization. (E to G) Representative FACS plots (E), percentage (F), and total number (G) of MERS-CoV-specific CD8 T cells in the lungs at 4 days after MERS MA 6.1.2 challenge ( n = 6 mice per group). Data are representative of two independent experiments. Data presented represent mean ± SE; * denotes P
    Figure Legend Snippet: Single-dose intranasal immunization with PIV5-MERS-S induced robust MERS-CoV-specific CD8 T cell response in human DPP4 knockin (hDPP4 KI) mice. (A) Schematic diagram showing the experimental plan to examine CD8 T cell response after immunization and challenge. hDPP4 KI mice were intranasally immunized with 10 4 PFU PIV5-MERS-S or PIV5-GFP. At 4 weeks or 4 days following immunization, mice were challenged with MERS MA 6.1.2, single-cell suspensions from the lungs of immunized mice were stimulated with MERS-CoV spike peptides (S343 and S1165), and specific CD8 T cells were determined by IFN-γ intracellular staining. (B to D) Representative FACS plots (B), percentage (C), and total number (D) of MERS-CoV-specific CD8 T cells in the lungs at 4 weeks after immunization. (E to G) Representative FACS plots (E), percentage (F), and total number (G) of MERS-CoV-specific CD8 T cells in the lungs at 4 days after MERS MA 6.1.2 challenge ( n = 6 mice per group). Data are representative of two independent experiments. Data presented represent mean ± SE; * denotes P

    Techniques Used: Knock-In, Mouse Assay, Staining, FACS

    Serum neutralizing antibodies produced in mice 4 weeks after single-dose intranasal immunization with PIV5-MERS-S. Naive mice were intranasally immunized with PIV5-GFP or PIV5-MERS-S. Sera were collected at 4 weeks postimmunization. Neutralization assay against MERS-CoV spike pseudovirions was performed as described in Materials and Methods. The neutralization results were measured in luciferase activity and plotted relative to mock-treatment value. (A and B) Neutralization assay results from C57BL/6 mice immunized with 10 4 PFU (A) and 10 6 PFU (B) PIV5-MERS-S or PIV5-GFP. (C) Neutralization assay results from BALB/c mice immunized with 10 6 PFU PIV5-MERS-S or PIV5-GFP. Data presented represent means ± SEs.
    Figure Legend Snippet: Serum neutralizing antibodies produced in mice 4 weeks after single-dose intranasal immunization with PIV5-MERS-S. Naive mice were intranasally immunized with PIV5-GFP or PIV5-MERS-S. Sera were collected at 4 weeks postimmunization. Neutralization assay against MERS-CoV spike pseudovirions was performed as described in Materials and Methods. The neutralization results were measured in luciferase activity and plotted relative to mock-treatment value. (A and B) Neutralization assay results from C57BL/6 mice immunized with 10 4 PFU (A) and 10 6 PFU (B) PIV5-MERS-S or PIV5-GFP. (C) Neutralization assay results from BALB/c mice immunized with 10 6 PFU PIV5-MERS-S or PIV5-GFP. Data presented represent means ± SEs.

    Techniques Used: Produced, Mouse Assay, Neutralization, Luciferase, Activity Assay

    Representative images of lung tissues from mice receiving PBS (A), UV-inactivated MERS-CoV (B), or PIV5-MERS-S (C) treatment, followed by infection with MERS MA 6.1.2. Images obtained from tissues at 3 days after MERS MA 6.1.2 infection. Compared to PBS or PIV5-MERS, perivascular eosinophilic infiltration (arrows) in UV-MERS-treated mice was greatly increased. n = 3 to 4 mice/group. (D) Graph representing eosinophil infiltration in the lung tissues of mice from groups shown in panels A to . * denotes P
    Figure Legend Snippet: Representative images of lung tissues from mice receiving PBS (A), UV-inactivated MERS-CoV (B), or PIV5-MERS-S (C) treatment, followed by infection with MERS MA 6.1.2. Images obtained from tissues at 3 days after MERS MA 6.1.2 infection. Compared to PBS or PIV5-MERS, perivascular eosinophilic infiltration (arrows) in UV-MERS-treated mice was greatly increased. n = 3 to 4 mice/group. (D) Graph representing eosinophil infiltration in the lung tissues of mice from groups shown in panels A to . * denotes P

    Techniques Used: Mouse Assay, Infection

    Histopathology in immunized mice challenged with MERS-CoV. hDPP4 KI mice were intranasally immunized with 10 4 PFU PIV5-MERS-S or PIV5-GFP. Four weeks after immunization, the mice were intranasally infected with 10 5 PFU MERS MA 6.1.2. (A) Representative images of H E-stained lung sections from PIV5-MERS-S- or PIV5-GFP-immunized hDPP4 KI mice at indicated days after MERS MA 6.1.2 challenge. Note the cellular infiltration (black arrows) and the hyaline membranes (red arrows). (B) Summary scores for disease in the lung sections. n = 3 to 5 mice/group. * denotes P
    Figure Legend Snippet: Histopathology in immunized mice challenged with MERS-CoV. hDPP4 KI mice were intranasally immunized with 10 4 PFU PIV5-MERS-S or PIV5-GFP. Four weeks after immunization, the mice were intranasally infected with 10 5 PFU MERS MA 6.1.2. (A) Representative images of H E-stained lung sections from PIV5-MERS-S- or PIV5-GFP-immunized hDPP4 KI mice at indicated days after MERS MA 6.1.2 challenge. Note the cellular infiltration (black arrows) and the hyaline membranes (red arrows). (B) Summary scores for disease in the lung sections. n = 3 to 5 mice/group. * denotes P

    Techniques Used: Histopathology, Mouse Assay, Infection, Staining

    Comparison of the protective efficacy between single-dose immunization with UV light-inactivated MERS-CoV and PIV5-MERS-S. hDPP4 KI mice were immunized with 10 4 PFU PIV5-MERS-S via intranasal route; 10 6 PFU UV-inactivated MERS MA 6.1.2, mixed with Imject alum; or PBS via intramuscular route. Four weeks after immunization, immunized mice were infected with 10 5 PFU MERS-CoV. (A) Schematic timeline outlining experimental plan. (B and C) Survival (B) and weight loss (C) were monitored daily until 10 days postinfection. PBS, n = 9; UV MERS-CoV, n = 12; PIV5-MERS-S, n = 8. Data represent mean ± SE.
    Figure Legend Snippet: Comparison of the protective efficacy between single-dose immunization with UV light-inactivated MERS-CoV and PIV5-MERS-S. hDPP4 KI mice were immunized with 10 4 PFU PIV5-MERS-S via intranasal route; 10 6 PFU UV-inactivated MERS MA 6.1.2, mixed with Imject alum; or PBS via intramuscular route. Four weeks after immunization, immunized mice were infected with 10 5 PFU MERS-CoV. (A) Schematic timeline outlining experimental plan. (B and C) Survival (B) and weight loss (C) were monitored daily until 10 days postinfection. PBS, n = 9; UV MERS-CoV, n = 12; PIV5-MERS-S, n = 8. Data represent mean ± SE.

    Techniques Used: Mouse Assay, Infection

    Generation and characterization of recombinant PIV5 expressing MERS-CoV spike protein. (A) Schematic of PIV5-MERS-S. NP, nucleoprotein; V, V protein; P, phosphoprotein; M, matrix protein; F, fusion protein; SH, small hydrophobic protein; HN, hemagglutinin-neuraminidase protein; L, RNA-dependent RNA polymerase. (B) Confirmation of MERS-CoV spike protein expression by Western blotting. Vero 81 cells were infected with PIV5-MERS-S at MOIs of 0.01, 0.1, and 1.0 or mock infected. At 2 days postinfection, MERS-CoV spike was detected with anti-MERS-S antibody by Western blotting. (C) Immunofluorescence of Vero cells infected with PIV5 and PIV5-MERS-S. Vero cells were infected with PIV5 and PIV5-MERS-S (MOI = 0.1) or mock infected. At 2 days postinfection, cells were fixed, permeabilized, and stained with anti-PIV5 V/P or anti-MERS-spike antibodies. Scale bar = 200 μm. (D) Growth rate of PIV5-MERS-S. MDBK cells were infected with PIV5 or PIV5-MERS-S at an MOI of 0.1. Media were collected daily for 5 days, and titers of viruses in the media were determined using plaque assay.
    Figure Legend Snippet: Generation and characterization of recombinant PIV5 expressing MERS-CoV spike protein. (A) Schematic of PIV5-MERS-S. NP, nucleoprotein; V, V protein; P, phosphoprotein; M, matrix protein; F, fusion protein; SH, small hydrophobic protein; HN, hemagglutinin-neuraminidase protein; L, RNA-dependent RNA polymerase. (B) Confirmation of MERS-CoV spike protein expression by Western blotting. Vero 81 cells were infected with PIV5-MERS-S at MOIs of 0.01, 0.1, and 1.0 or mock infected. At 2 days postinfection, MERS-CoV spike was detected with anti-MERS-S antibody by Western blotting. (C) Immunofluorescence of Vero cells infected with PIV5 and PIV5-MERS-S. Vero cells were infected with PIV5 and PIV5-MERS-S (MOI = 0.1) or mock infected. At 2 days postinfection, cells were fixed, permeabilized, and stained with anti-PIV5 V/P or anti-MERS-spike antibodies. Scale bar = 200 μm. (D) Growth rate of PIV5-MERS-S. MDBK cells were infected with PIV5 or PIV5-MERS-S at an MOI of 0.1. Media were collected daily for 5 days, and titers of viruses in the media were determined using plaque assay.

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

    Single-dose intranasal immunization with PIV5-MERS-S completely protects hDPP4 KI mice from lethal MERS-CoV challenge. (A) Schematic timeline showing immunization, challenge, and the evaluation of protection. hDPP4 KI mice were intranasally immunized with 10 4 PFU PIV5-MERS-S, PIV5-GFP, or PBS. Four weeks after immunization, the mice were intranasally infected with 10 5 PFU MERS MA 6.1.2. (B and C) Survival (B) and weight loss (C) were monitored daily for 12 days. PIV5-MERS-S or PIV5-GFP, n = 10; PBS, n = 5. (D) At indicated days postinfection, virus lung titers were quantified by plaque assay. Data are representative of two independent experiments. Data presented represent mean ± SE; * denotes P
    Figure Legend Snippet: Single-dose intranasal immunization with PIV5-MERS-S completely protects hDPP4 KI mice from lethal MERS-CoV challenge. (A) Schematic timeline showing immunization, challenge, and the evaluation of protection. hDPP4 KI mice were intranasally immunized with 10 4 PFU PIV5-MERS-S, PIV5-GFP, or PBS. Four weeks after immunization, the mice were intranasally infected with 10 5 PFU MERS MA 6.1.2. (B and C) Survival (B) and weight loss (C) were monitored daily for 12 days. PIV5-MERS-S or PIV5-GFP, n = 10; PBS, n = 5. (D) At indicated days postinfection, virus lung titers were quantified by plaque assay. Data are representative of two independent experiments. Data presented represent mean ± SE; * denotes P

    Techniques Used: Mouse Assay, Infection, Plaque Assay

    Related Articles

    Expressing:

    Article Title: Inhibition of Proprotein Convertases Abrogates Processing of the Middle Eastern Respiratory Syndrome Coronavirus Spike Protein in Infected Cells but Does Not Reduce Viral Infectivity
    Article Snippet: At 24 hours after infection, the cells were washed and harvested, and the pellet was lysed with RIPA lysis buffer, supplemented with 4xNuPAGE (Invitrogen) and boiled for 20 minutes at 95°C. .. S protein expression in lysates was detected by Western blot, using a polyclonal antibody directed against the S2 subunit of MERS-S (Sino Biological). .. In parallel, for quantification of viral RNA, 50 µL of the cell supernatant was dissolved in RAV1 buffer (Macherey-Nagel) for RNA extraction, followed by quantitative reverse-transcription PCR analysis, using the upE assay as previously described [ ].

    Western Blot:

    Article Title: Inhibition of Proprotein Convertases Abrogates Processing of the Middle Eastern Respiratory Syndrome Coronavirus Spike Protein in Infected Cells but Does Not Reduce Viral Infectivity
    Article Snippet: At 24 hours after infection, the cells were washed and harvested, and the pellet was lysed with RIPA lysis buffer, supplemented with 4xNuPAGE (Invitrogen) and boiled for 20 minutes at 95°C. .. S protein expression in lysates was detected by Western blot, using a polyclonal antibody directed against the S2 subunit of MERS-S (Sino Biological). .. In parallel, for quantification of viral RNA, 50 µL of the cell supernatant was dissolved in RAV1 buffer (Macherey-Nagel) for RNA extraction, followed by quantitative reverse-transcription PCR analysis, using the upE assay as previously described [ ].

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  • 93
    Sino Biological mers s
    RXXR motifs located at the border between S1 and <t>S2</t> are required for efficient processing of the Middle East respiratory syndrome coronavirus spike protein <t>(MERS-S).</t> A , The domain organization of the MERS-S protein is schematically depicted. The MERS-S sequence at the border between the S1 and S2 subunits is shown. RXXR motifs, which constitute potential cleavage sites, are highlighted, and the predicted start of the S2 subunit is underlined. The mutations introduced into the potential cleavage sites in MERS-S are shown. B , 293T cells were transfected with expression plasmids coding for MERS-S wild type and the indicated MERS-S mutants equipped with a C-terminal V5 tag. Transfection of empty plasmid (pcDNA) served as negative control. Expression of S proteins in cell lysates was determined by Western blot, using a V5 tag–specific monoclonal antibody. Expression of β-actin in cell lysates was assessed as a loading control. The results shown are representative for at least 3 independent experiments. Abbreviations: CT, cytoplasmic tail; PCM, potential cleavage site mutant; RBD, receptor binding domain; SP, signal peptide; TM, transmembrane domain.
    Mers S, supplied by Sino Biological, 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/mers s/product/Sino Biological
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mers s - by Bioz Stars, 2021-06
    93/100 stars
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    RXXR motifs located at the border between S1 and S2 are required for efficient processing of the Middle East respiratory syndrome coronavirus spike protein (MERS-S). A , The domain organization of the MERS-S protein is schematically depicted. The MERS-S sequence at the border between the S1 and S2 subunits is shown. RXXR motifs, which constitute potential cleavage sites, are highlighted, and the predicted start of the S2 subunit is underlined. The mutations introduced into the potential cleavage sites in MERS-S are shown. B , 293T cells were transfected with expression plasmids coding for MERS-S wild type and the indicated MERS-S mutants equipped with a C-terminal V5 tag. Transfection of empty plasmid (pcDNA) served as negative control. Expression of S proteins in cell lysates was determined by Western blot, using a V5 tag–specific monoclonal antibody. Expression of β-actin in cell lysates was assessed as a loading control. The results shown are representative for at least 3 independent experiments. Abbreviations: CT, cytoplasmic tail; PCM, potential cleavage site mutant; RBD, receptor binding domain; SP, signal peptide; TM, transmembrane domain.

    Journal: The Journal of Infectious Diseases

    Article Title: Inhibition of Proprotein Convertases Abrogates Processing of the Middle Eastern Respiratory Syndrome Coronavirus Spike Protein in Infected Cells but Does Not Reduce Viral Infectivity

    doi: 10.1093/infdis/jiu407

    Figure Lengend Snippet: RXXR motifs located at the border between S1 and S2 are required for efficient processing of the Middle East respiratory syndrome coronavirus spike protein (MERS-S). A , The domain organization of the MERS-S protein is schematically depicted. The MERS-S sequence at the border between the S1 and S2 subunits is shown. RXXR motifs, which constitute potential cleavage sites, are highlighted, and the predicted start of the S2 subunit is underlined. The mutations introduced into the potential cleavage sites in MERS-S are shown. B , 293T cells were transfected with expression plasmids coding for MERS-S wild type and the indicated MERS-S mutants equipped with a C-terminal V5 tag. Transfection of empty plasmid (pcDNA) served as negative control. Expression of S proteins in cell lysates was determined by Western blot, using a V5 tag–specific monoclonal antibody. Expression of β-actin in cell lysates was assessed as a loading control. The results shown are representative for at least 3 independent experiments. Abbreviations: CT, cytoplasmic tail; PCM, potential cleavage site mutant; RBD, receptor binding domain; SP, signal peptide; TM, transmembrane domain.

    Article Snippet: S protein expression in lysates was detected by Western blot, using a polyclonal antibody directed against the S2 subunit of MERS-S (Sino Biological).

    Techniques: Sequencing, Transfection, Expressing, Plasmid Preparation, Negative Control, Western Blot, Mutagenesis, Binding Assay

    Proprotein convertase activity is dispensable for Middle East respiratory syndrome coronavirus spike protein (MERS-S)–driven cell-cell and virus-cell fusion. A , Fusion of 293T effector cells transfected to express MERS-S with target cells transfected to express DPP4 and/or TMPRSS2 or control transfected with empty plasmid (pcDNA) was assessed. Both effector and target cells were incubated with 1 µM of proprotein convertase inhibitor (PCI) as indicated and, 1 day later, were mixed for cocultivation. The effector/target cell mixtures were incubated with phosphate-buffered saline (PBS) or PCI, and cell-cell fusion was quantified by determination of luciferase activities in cell lysates. The results of a representative experiment performed with triplicate samples are shown. Error bars indicate standard deviation (SD). Two separate experiments yielded similar results. B , Lentiviral pseudotypes carrying MERS-S, Lassa virus glycoprotein (LASV-GPC), Zaire ebolavirus glycoprotein (EBOV-GP), or the glycoprotein of vesicular stomatitis virus (VSV-G), as well as pseudotypes bearing no viral glycoprotein (pcDNA), were generated in the presence or absence of PCI (1 µM). Subsequently, target 293T cells transfected with empty plasmid or DPP4 expression plasmid were preincubated with dimethyl sulfoxide (DMSO), PBS, or PCI at a final concentration of 0.5 µM for 30–60 minutes, followed by transduction with the pseudotypes specified above. At 72 hours after transduction, luciferase activities in cell lysates were measured. The average of 5–7 independent experiments performed with triplicate samples is shown. Transduction with pseudotypes bearing VSV-G in the absence of inhibitor was set as 100%. Error bars indicate standard error of the mean (SEM). A 2-tailed Student t test was used to assess statistical significance.

    Journal: The Journal of Infectious Diseases

    Article Title: Inhibition of Proprotein Convertases Abrogates Processing of the Middle Eastern Respiratory Syndrome Coronavirus Spike Protein in Infected Cells but Does Not Reduce Viral Infectivity

    doi: 10.1093/infdis/jiu407

    Figure Lengend Snippet: Proprotein convertase activity is dispensable for Middle East respiratory syndrome coronavirus spike protein (MERS-S)–driven cell-cell and virus-cell fusion. A , Fusion of 293T effector cells transfected to express MERS-S with target cells transfected to express DPP4 and/or TMPRSS2 or control transfected with empty plasmid (pcDNA) was assessed. Both effector and target cells were incubated with 1 µM of proprotein convertase inhibitor (PCI) as indicated and, 1 day later, were mixed for cocultivation. The effector/target cell mixtures were incubated with phosphate-buffered saline (PBS) or PCI, and cell-cell fusion was quantified by determination of luciferase activities in cell lysates. The results of a representative experiment performed with triplicate samples are shown. Error bars indicate standard deviation (SD). Two separate experiments yielded similar results. B , Lentiviral pseudotypes carrying MERS-S, Lassa virus glycoprotein (LASV-GPC), Zaire ebolavirus glycoprotein (EBOV-GP), or the glycoprotein of vesicular stomatitis virus (VSV-G), as well as pseudotypes bearing no viral glycoprotein (pcDNA), were generated in the presence or absence of PCI (1 µM). Subsequently, target 293T cells transfected with empty plasmid or DPP4 expression plasmid were preincubated with dimethyl sulfoxide (DMSO), PBS, or PCI at a final concentration of 0.5 µM for 30–60 minutes, followed by transduction with the pseudotypes specified above. At 72 hours after transduction, luciferase activities in cell lysates were measured. The average of 5–7 independent experiments performed with triplicate samples is shown. Transduction with pseudotypes bearing VSV-G in the absence of inhibitor was set as 100%. Error bars indicate standard error of the mean (SEM). A 2-tailed Student t test was used to assess statistical significance.

    Article Snippet: S protein expression in lysates was detected by Western blot, using a polyclonal antibody directed against the S2 subunit of MERS-S (Sino Biological).

    Techniques: Activity Assay, Transfection, Plasmid Preparation, Incubation, Luciferase, Standard Deviation, Gel Permeation Chromatography, Generated, Expressing, Concentration Assay, Transduction

    The Middle East respiratory syndrome coronavirus spike protein (MERS-S) is cleaved by proprotein convertases. 293T cells were transfected with expression plasmids encoding MERS-S, Zaire ebolavirus glycoprotein (EBOV-GP), or Lassa virus glycoprotein (LASV-GPC), all equipped with a C-terminal V5 tag. Cells transfected with empty plasmid (pcDNA) served as negative control. Subsequently, cells were incubated with the indicated concentrations of the proprotein convertase inhibitor (PCI). At 48 hours after transfection, glycoprotein expression was analyzed by Western blot, using a V5 tag–specific monoclonal antibody. Detection of β-actin served as loading control. The results shown are representative of three independent experiments.

    Journal: The Journal of Infectious Diseases

    Article Title: Inhibition of Proprotein Convertases Abrogates Processing of the Middle Eastern Respiratory Syndrome Coronavirus Spike Protein in Infected Cells but Does Not Reduce Viral Infectivity

    doi: 10.1093/infdis/jiu407

    Figure Lengend Snippet: The Middle East respiratory syndrome coronavirus spike protein (MERS-S) is cleaved by proprotein convertases. 293T cells were transfected with expression plasmids encoding MERS-S, Zaire ebolavirus glycoprotein (EBOV-GP), or Lassa virus glycoprotein (LASV-GPC), all equipped with a C-terminal V5 tag. Cells transfected with empty plasmid (pcDNA) served as negative control. Subsequently, cells were incubated with the indicated concentrations of the proprotein convertase inhibitor (PCI). At 48 hours after transfection, glycoprotein expression was analyzed by Western blot, using a V5 tag–specific monoclonal antibody. Detection of β-actin served as loading control. The results shown are representative of three independent experiments.

    Article Snippet: S protein expression in lysates was detected by Western blot, using a polyclonal antibody directed against the S2 subunit of MERS-S (Sino Biological).

    Techniques: Transfection, Expressing, Gel Permeation Chromatography, Plasmid Preparation, Negative Control, Incubation, Western Blot

    The Middle East respiratory syndrome coronavirus (MERS-CoV) spike protein (MERS-S) is cleaved in transfected and infected cells. 293T cells were transfected with a plasmid encoding the MERS-S protein or with empty plasmid (pcDNA). Vero B4 cells were either infected with MERS-CoV at a multiplicity of infection of 5 or mock infected. Subsequently, the cells were lysed and analyzed by Western blot, using a polyclonal antibody directed against the S2 subunit of MERS-S. A β-actin antibody served as a loading control. Similar results were obtained in 2 separate experiments.

    Journal: The Journal of Infectious Diseases

    Article Title: Inhibition of Proprotein Convertases Abrogates Processing of the Middle Eastern Respiratory Syndrome Coronavirus Spike Protein in Infected Cells but Does Not Reduce Viral Infectivity

    doi: 10.1093/infdis/jiu407

    Figure Lengend Snippet: The Middle East respiratory syndrome coronavirus (MERS-CoV) spike protein (MERS-S) is cleaved in transfected and infected cells. 293T cells were transfected with a plasmid encoding the MERS-S protein or with empty plasmid (pcDNA). Vero B4 cells were either infected with MERS-CoV at a multiplicity of infection of 5 or mock infected. Subsequently, the cells were lysed and analyzed by Western blot, using a polyclonal antibody directed against the S2 subunit of MERS-S. A β-actin antibody served as a loading control. Similar results were obtained in 2 separate experiments.

    Article Snippet: S protein expression in lysates was detected by Western blot, using a polyclonal antibody directed against the S2 subunit of MERS-S (Sino Biological).

    Techniques: Transfection, Infection, Plasmid Preparation, Western Blot

    Single-dose intranasal immunization with PIV5-MERS-S induced robust MERS-CoV-specific CD8 T cell response in human DPP4 knockin (hDPP4 KI) mice. (A) Schematic diagram showing the experimental plan to examine CD8 T cell response after immunization and challenge. hDPP4 KI mice were intranasally immunized with 10 4 PFU PIV5-MERS-S or PIV5-GFP. At 4 weeks or 4 days following immunization, mice were challenged with MERS MA 6.1.2, single-cell suspensions from the lungs of immunized mice were stimulated with MERS-CoV spike peptides (S343 and S1165), and specific CD8 T cells were determined by IFN-γ intracellular staining. (B to D) Representative FACS plots (B), percentage (C), and total number (D) of MERS-CoV-specific CD8 T cells in the lungs at 4 weeks after immunization. (E to G) Representative FACS plots (E), percentage (F), and total number (G) of MERS-CoV-specific CD8 T cells in the lungs at 4 days after MERS MA 6.1.2 challenge ( n = 6 mice per group). Data are representative of two independent experiments. Data presented represent mean ± SE; * denotes P

    Journal: mBio

    Article Title: Single-Dose, Intranasal Immunization with Recombinant Parainfluenza Virus 5 Expressing Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Spike Protein Protects Mice from Fatal MERS-CoV Infection

    doi: 10.1128/mBio.00554-20

    Figure Lengend Snippet: Single-dose intranasal immunization with PIV5-MERS-S induced robust MERS-CoV-specific CD8 T cell response in human DPP4 knockin (hDPP4 KI) mice. (A) Schematic diagram showing the experimental plan to examine CD8 T cell response after immunization and challenge. hDPP4 KI mice were intranasally immunized with 10 4 PFU PIV5-MERS-S or PIV5-GFP. At 4 weeks or 4 days following immunization, mice were challenged with MERS MA 6.1.2, single-cell suspensions from the lungs of immunized mice were stimulated with MERS-CoV spike peptides (S343 and S1165), and specific CD8 T cells were determined by IFN-γ intracellular staining. (B to D) Representative FACS plots (B), percentage (C), and total number (D) of MERS-CoV-specific CD8 T cells in the lungs at 4 weeks after immunization. (E to G) Representative FACS plots (E), percentage (F), and total number (G) of MERS-CoV-specific CD8 T cells in the lungs at 4 days after MERS MA 6.1.2 challenge ( n = 6 mice per group). Data are representative of two independent experiments. Data presented represent mean ± SE; * denotes P

    Article Snippet: Anti-MERS-S from Sino Biological was used (catalog no. 40070-T60).

    Techniques: Knock-In, Mouse Assay, Staining, FACS

    Serum neutralizing antibodies produced in mice 4 weeks after single-dose intranasal immunization with PIV5-MERS-S. Naive mice were intranasally immunized with PIV5-GFP or PIV5-MERS-S. Sera were collected at 4 weeks postimmunization. Neutralization assay against MERS-CoV spike pseudovirions was performed as described in Materials and Methods. The neutralization results were measured in luciferase activity and plotted relative to mock-treatment value. (A and B) Neutralization assay results from C57BL/6 mice immunized with 10 4 PFU (A) and 10 6 PFU (B) PIV5-MERS-S or PIV5-GFP. (C) Neutralization assay results from BALB/c mice immunized with 10 6 PFU PIV5-MERS-S or PIV5-GFP. Data presented represent means ± SEs.

    Journal: mBio

    Article Title: Single-Dose, Intranasal Immunization with Recombinant Parainfluenza Virus 5 Expressing Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Spike Protein Protects Mice from Fatal MERS-CoV Infection

    doi: 10.1128/mBio.00554-20

    Figure Lengend Snippet: Serum neutralizing antibodies produced in mice 4 weeks after single-dose intranasal immunization with PIV5-MERS-S. Naive mice were intranasally immunized with PIV5-GFP or PIV5-MERS-S. Sera were collected at 4 weeks postimmunization. Neutralization assay against MERS-CoV spike pseudovirions was performed as described in Materials and Methods. The neutralization results were measured in luciferase activity and plotted relative to mock-treatment value. (A and B) Neutralization assay results from C57BL/6 mice immunized with 10 4 PFU (A) and 10 6 PFU (B) PIV5-MERS-S or PIV5-GFP. (C) Neutralization assay results from BALB/c mice immunized with 10 6 PFU PIV5-MERS-S or PIV5-GFP. Data presented represent means ± SEs.

    Article Snippet: Anti-MERS-S from Sino Biological was used (catalog no. 40070-T60).

    Techniques: Produced, Mouse Assay, Neutralization, Luciferase, Activity Assay

    Representative images of lung tissues from mice receiving PBS (A), UV-inactivated MERS-CoV (B), or PIV5-MERS-S (C) treatment, followed by infection with MERS MA 6.1.2. Images obtained from tissues at 3 days after MERS MA 6.1.2 infection. Compared to PBS or PIV5-MERS, perivascular eosinophilic infiltration (arrows) in UV-MERS-treated mice was greatly increased. n = 3 to 4 mice/group. (D) Graph representing eosinophil infiltration in the lung tissues of mice from groups shown in panels A to . * denotes P

    Journal: mBio

    Article Title: Single-Dose, Intranasal Immunization with Recombinant Parainfluenza Virus 5 Expressing Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Spike Protein Protects Mice from Fatal MERS-CoV Infection

    doi: 10.1128/mBio.00554-20

    Figure Lengend Snippet: Representative images of lung tissues from mice receiving PBS (A), UV-inactivated MERS-CoV (B), or PIV5-MERS-S (C) treatment, followed by infection with MERS MA 6.1.2. Images obtained from tissues at 3 days after MERS MA 6.1.2 infection. Compared to PBS or PIV5-MERS, perivascular eosinophilic infiltration (arrows) in UV-MERS-treated mice was greatly increased. n = 3 to 4 mice/group. (D) Graph representing eosinophil infiltration in the lung tissues of mice from groups shown in panels A to . * denotes P

    Article Snippet: Anti-MERS-S from Sino Biological was used (catalog no. 40070-T60).

    Techniques: Mouse Assay, Infection

    Histopathology in immunized mice challenged with MERS-CoV. hDPP4 KI mice were intranasally immunized with 10 4 PFU PIV5-MERS-S or PIV5-GFP. Four weeks after immunization, the mice were intranasally infected with 10 5 PFU MERS MA 6.1.2. (A) Representative images of H E-stained lung sections from PIV5-MERS-S- or PIV5-GFP-immunized hDPP4 KI mice at indicated days after MERS MA 6.1.2 challenge. Note the cellular infiltration (black arrows) and the hyaline membranes (red arrows). (B) Summary scores for disease in the lung sections. n = 3 to 5 mice/group. * denotes P

    Journal: mBio

    Article Title: Single-Dose, Intranasal Immunization with Recombinant Parainfluenza Virus 5 Expressing Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Spike Protein Protects Mice from Fatal MERS-CoV Infection

    doi: 10.1128/mBio.00554-20

    Figure Lengend Snippet: Histopathology in immunized mice challenged with MERS-CoV. hDPP4 KI mice were intranasally immunized with 10 4 PFU PIV5-MERS-S or PIV5-GFP. Four weeks after immunization, the mice were intranasally infected with 10 5 PFU MERS MA 6.1.2. (A) Representative images of H E-stained lung sections from PIV5-MERS-S- or PIV5-GFP-immunized hDPP4 KI mice at indicated days after MERS MA 6.1.2 challenge. Note the cellular infiltration (black arrows) and the hyaline membranes (red arrows). (B) Summary scores for disease in the lung sections. n = 3 to 5 mice/group. * denotes P

    Article Snippet: Anti-MERS-S from Sino Biological was used (catalog no. 40070-T60).

    Techniques: Histopathology, Mouse Assay, Infection, Staining

    Comparison of the protective efficacy between single-dose immunization with UV light-inactivated MERS-CoV and PIV5-MERS-S. hDPP4 KI mice were immunized with 10 4 PFU PIV5-MERS-S via intranasal route; 10 6 PFU UV-inactivated MERS MA 6.1.2, mixed with Imject alum; or PBS via intramuscular route. Four weeks after immunization, immunized mice were infected with 10 5 PFU MERS-CoV. (A) Schematic timeline outlining experimental plan. (B and C) Survival (B) and weight loss (C) were monitored daily until 10 days postinfection. PBS, n = 9; UV MERS-CoV, n = 12; PIV5-MERS-S, n = 8. Data represent mean ± SE.

    Journal: mBio

    Article Title: Single-Dose, Intranasal Immunization with Recombinant Parainfluenza Virus 5 Expressing Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Spike Protein Protects Mice from Fatal MERS-CoV Infection

    doi: 10.1128/mBio.00554-20

    Figure Lengend Snippet: Comparison of the protective efficacy between single-dose immunization with UV light-inactivated MERS-CoV and PIV5-MERS-S. hDPP4 KI mice were immunized with 10 4 PFU PIV5-MERS-S via intranasal route; 10 6 PFU UV-inactivated MERS MA 6.1.2, mixed with Imject alum; or PBS via intramuscular route. Four weeks after immunization, immunized mice were infected with 10 5 PFU MERS-CoV. (A) Schematic timeline outlining experimental plan. (B and C) Survival (B) and weight loss (C) were monitored daily until 10 days postinfection. PBS, n = 9; UV MERS-CoV, n = 12; PIV5-MERS-S, n = 8. Data represent mean ± SE.

    Article Snippet: Anti-MERS-S from Sino Biological was used (catalog no. 40070-T60).

    Techniques: Mouse Assay, Infection

    Generation and characterization of recombinant PIV5 expressing MERS-CoV spike protein. (A) Schematic of PIV5-MERS-S. NP, nucleoprotein; V, V protein; P, phosphoprotein; M, matrix protein; F, fusion protein; SH, small hydrophobic protein; HN, hemagglutinin-neuraminidase protein; L, RNA-dependent RNA polymerase. (B) Confirmation of MERS-CoV spike protein expression by Western blotting. Vero 81 cells were infected with PIV5-MERS-S at MOIs of 0.01, 0.1, and 1.0 or mock infected. At 2 days postinfection, MERS-CoV spike was detected with anti-MERS-S antibody by Western blotting. (C) Immunofluorescence of Vero cells infected with PIV5 and PIV5-MERS-S. Vero cells were infected with PIV5 and PIV5-MERS-S (MOI = 0.1) or mock infected. At 2 days postinfection, cells were fixed, permeabilized, and stained with anti-PIV5 V/P or anti-MERS-spike antibodies. Scale bar = 200 μm. (D) Growth rate of PIV5-MERS-S. MDBK cells were infected with PIV5 or PIV5-MERS-S at an MOI of 0.1. Media were collected daily for 5 days, and titers of viruses in the media were determined using plaque assay.

    Journal: mBio

    Article Title: Single-Dose, Intranasal Immunization with Recombinant Parainfluenza Virus 5 Expressing Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Spike Protein Protects Mice from Fatal MERS-CoV Infection

    doi: 10.1128/mBio.00554-20

    Figure Lengend Snippet: Generation and characterization of recombinant PIV5 expressing MERS-CoV spike protein. (A) Schematic of PIV5-MERS-S. NP, nucleoprotein; V, V protein; P, phosphoprotein; M, matrix protein; F, fusion protein; SH, small hydrophobic protein; HN, hemagglutinin-neuraminidase protein; L, RNA-dependent RNA polymerase. (B) Confirmation of MERS-CoV spike protein expression by Western blotting. Vero 81 cells were infected with PIV5-MERS-S at MOIs of 0.01, 0.1, and 1.0 or mock infected. At 2 days postinfection, MERS-CoV spike was detected with anti-MERS-S antibody by Western blotting. (C) Immunofluorescence of Vero cells infected with PIV5 and PIV5-MERS-S. Vero cells were infected with PIV5 and PIV5-MERS-S (MOI = 0.1) or mock infected. At 2 days postinfection, cells were fixed, permeabilized, and stained with anti-PIV5 V/P or anti-MERS-spike antibodies. Scale bar = 200 μm. (D) Growth rate of PIV5-MERS-S. MDBK cells were infected with PIV5 or PIV5-MERS-S at an MOI of 0.1. Media were collected daily for 5 days, and titers of viruses in the media were determined using plaque assay.

    Article Snippet: Anti-MERS-S from Sino Biological was used (catalog no. 40070-T60).

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

    Single-dose intranasal immunization with PIV5-MERS-S completely protects hDPP4 KI mice from lethal MERS-CoV challenge. (A) Schematic timeline showing immunization, challenge, and the evaluation of protection. hDPP4 KI mice were intranasally immunized with 10 4 PFU PIV5-MERS-S, PIV5-GFP, or PBS. Four weeks after immunization, the mice were intranasally infected with 10 5 PFU MERS MA 6.1.2. (B and C) Survival (B) and weight loss (C) were monitored daily for 12 days. PIV5-MERS-S or PIV5-GFP, n = 10; PBS, n = 5. (D) At indicated days postinfection, virus lung titers were quantified by plaque assay. Data are representative of two independent experiments. Data presented represent mean ± SE; * denotes P

    Journal: mBio

    Article Title: Single-Dose, Intranasal Immunization with Recombinant Parainfluenza Virus 5 Expressing Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Spike Protein Protects Mice from Fatal MERS-CoV Infection

    doi: 10.1128/mBio.00554-20

    Figure Lengend Snippet: Single-dose intranasal immunization with PIV5-MERS-S completely protects hDPP4 KI mice from lethal MERS-CoV challenge. (A) Schematic timeline showing immunization, challenge, and the evaluation of protection. hDPP4 KI mice were intranasally immunized with 10 4 PFU PIV5-MERS-S, PIV5-GFP, or PBS. Four weeks after immunization, the mice were intranasally infected with 10 5 PFU MERS MA 6.1.2. (B and C) Survival (B) and weight loss (C) were monitored daily for 12 days. PIV5-MERS-S or PIV5-GFP, n = 10; PBS, n = 5. (D) At indicated days postinfection, virus lung titers were quantified by plaque assay. Data are representative of two independent experiments. Data presented represent mean ± SE; * denotes P

    Article Snippet: Anti-MERS-S from Sino Biological was used (catalog no. 40070-T60).

    Techniques: Mouse Assay, Infection, Plaque Assay