vero 76 cells  (ATCC)


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    Name:
    VERO 76
    Description:

    Catalog Number:
    crl-1587
    Price:
    None
    Host:
    Cercopithecus aethiops
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    Structured Review

    ATCC vero 76 cells
    JUNV replication in CD34 + cells. CD34 + cells were inoculated with UV-irradiated JUNV or JUNV and stimulated with TPO for ten days. Viral replication was assayed by RT-PCR, immunofluorescence and flow cytometry. (A) RT-PCR studies. (B) To detect JUNV antigens by immunofluorescence cells were washed, cytocentrifuged on silanized glasses, fixed, permeabilized and incubated first with a pool of specific mAbs against JUNV and a rabbit-anti-human vWF polyclonal Ab to identify megakaryocytes, and then with FITC-conjugated anti-rabbit (green) and Cy3-conjugated anti-mouse Igs (red). The slides were counterstained with DAPI and photographed at 1000× magnification. (C) Cells were stained as in B and then analyzed by flow cytometry. (D) As a positive control, JUNV-susceptible <t>Vero-76</t> cells were inoculated with JUNV and seven days later were stained with the pool of specific mAbs against JUNV followed by Cy3-anti-mouse Igs. Negative controls in B and C were performed by incubating cells only with secondary Abs. Panels show a representative experiment of three similar replicates.

    https://www.bioz.com/result/vero 76 cells/product/ATCC
    Average 99 stars, based on 21 article reviews
    Price from $9.99 to $1999.99
    vero 76 cells - by Bioz Stars, 2020-11
    99/100 stars

    Images

    1) Product Images from "Jun?n Virus Infection of Human Hematopoietic Progenitors Impairs In Vitro Proplatelet Formation and Platelet Release via a Bystander Effect Involving Type I IFN Signaling"

    Article Title: Jun?n Virus Infection of Human Hematopoietic Progenitors Impairs In Vitro Proplatelet Formation and Platelet Release via a Bystander Effect Involving Type I IFN Signaling

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1000847

    JUNV replication in CD34 + cells. CD34 + cells were inoculated with UV-irradiated JUNV or JUNV and stimulated with TPO for ten days. Viral replication was assayed by RT-PCR, immunofluorescence and flow cytometry. (A) RT-PCR studies. (B) To detect JUNV antigens by immunofluorescence cells were washed, cytocentrifuged on silanized glasses, fixed, permeabilized and incubated first with a pool of specific mAbs against JUNV and a rabbit-anti-human vWF polyclonal Ab to identify megakaryocytes, and then with FITC-conjugated anti-rabbit (green) and Cy3-conjugated anti-mouse Igs (red). The slides were counterstained with DAPI and photographed at 1000× magnification. (C) Cells were stained as in B and then analyzed by flow cytometry. (D) As a positive control, JUNV-susceptible Vero-76 cells were inoculated with JUNV and seven days later were stained with the pool of specific mAbs against JUNV followed by Cy3-anti-mouse Igs. Negative controls in B and C were performed by incubating cells only with secondary Abs. Panels show a representative experiment of three similar replicates.
    Figure Legend Snippet: JUNV replication in CD34 + cells. CD34 + cells were inoculated with UV-irradiated JUNV or JUNV and stimulated with TPO for ten days. Viral replication was assayed by RT-PCR, immunofluorescence and flow cytometry. (A) RT-PCR studies. (B) To detect JUNV antigens by immunofluorescence cells were washed, cytocentrifuged on silanized glasses, fixed, permeabilized and incubated first with a pool of specific mAbs against JUNV and a rabbit-anti-human vWF polyclonal Ab to identify megakaryocytes, and then with FITC-conjugated anti-rabbit (green) and Cy3-conjugated anti-mouse Igs (red). The slides were counterstained with DAPI and photographed at 1000× magnification. (C) Cells were stained as in B and then analyzed by flow cytometry. (D) As a positive control, JUNV-susceptible Vero-76 cells were inoculated with JUNV and seven days later were stained with the pool of specific mAbs against JUNV followed by Cy3-anti-mouse Igs. Negative controls in B and C were performed by incubating cells only with secondary Abs. Panels show a representative experiment of three similar replicates.

    Techniques Used: Irradiation, Reverse Transcription Polymerase Chain Reaction, Immunofluorescence, Flow Cytometry, Cytometry, Incubation, Staining, Positive Control

    2) Product Images from "Epistastic Interactions within the Junín Virus Envelope Glycoprotein Complex Provide an Evolutionary Barrier to Reversion in the Live-Attenuated Candid#1 Vaccine"

    Article Title: Epistastic Interactions within the Junín Virus Envelope Glycoprotein Complex Provide an Evolutionary Barrier to Reversion in the Live-Attenuated Candid#1 Vaccine

    Journal: Journal of Virology

    doi: 10.1128/JVI.01682-17

    Effect of A168T and F427I mutations on expression and cell-cell fusion activity of ectopically expressed GPC. (A and B) Metabolically labeled Candid#1 and MC2 GPCs were immunoprecipitated from transfected Vero-76 cells using the anti-GP1 MAb BF11 and resolved using NuPAGE. The mature GP1 and GP2 subunits comigrate as glycoproteins (A) and are resolved following deglycosylation by treatment with PNGase F (B). The uncleaved GP1GP2 precursor is labeled, and deglycosylated polypeptides are indicated as pp. GPC expression was quantitated by using a Fuji FLA3000G phosphorimager and ImageGauge software. Photostimulated luminescence (PSL) values in the respective GP1GP2 precursor bands in panel A are 427,881, 174,626, 360,045, 68,087, 339,735, and 53,852. This pattern of expression was confirmed in three replicate analyses. (C) Cell-cell fusion activity of the ectopically expressed MC2 (red) and Candid#1 (blue) GPCs was triggered by exposure to medium adjusted to pH 5.0 and determined using a chemiluminescent β-galactosidase fusion reporter. Relative light units are plotted. Error bars represent the standard errors of the means from replicate fusion reactions. Complete and partial replicates of this experiment ( n > 4) yielded concordant results. A pairwise statistical comparison using a 2-tailed Mann-Whitney analysis, as implemented in GraphPad Prism software, demonstrated that Can GPC is significantly more fusogenic than MC2 GPC ( P = 0.0002).
    Figure Legend Snippet: Effect of A168T and F427I mutations on expression and cell-cell fusion activity of ectopically expressed GPC. (A and B) Metabolically labeled Candid#1 and MC2 GPCs were immunoprecipitated from transfected Vero-76 cells using the anti-GP1 MAb BF11 and resolved using NuPAGE. The mature GP1 and GP2 subunits comigrate as glycoproteins (A) and are resolved following deglycosylation by treatment with PNGase F (B). The uncleaved GP1GP2 precursor is labeled, and deglycosylated polypeptides are indicated as pp. GPC expression was quantitated by using a Fuji FLA3000G phosphorimager and ImageGauge software. Photostimulated luminescence (PSL) values in the respective GP1GP2 precursor bands in panel A are 427,881, 174,626, 360,045, 68,087, 339,735, and 53,852. This pattern of expression was confirmed in three replicate analyses. (C) Cell-cell fusion activity of the ectopically expressed MC2 (red) and Candid#1 (blue) GPCs was triggered by exposure to medium adjusted to pH 5.0 and determined using a chemiluminescent β-galactosidase fusion reporter. Relative light units are plotted. Error bars represent the standard errors of the means from replicate fusion reactions. Complete and partial replicates of this experiment ( n > 4) yielded concordant results. A pairwise statistical comparison using a 2-tailed Mann-Whitney analysis, as implemented in GraphPad Prism software, demonstrated that Can GPC is significantly more fusogenic than MC2 GPC ( P = 0.0002).

    Techniques Used: Expressing, Activity Assay, Gel Permeation Chromatography, Metabolic Labelling, Labeling, Immunoprecipitation, Transfection, Software, MANN-WHITNEY

    3) Product Images from "Jun?n Virus Infection of Human Hematopoietic Progenitors Impairs In Vitro Proplatelet Formation and Platelet Release via a Bystander Effect Involving Type I IFN Signaling"

    Article Title: Jun?n Virus Infection of Human Hematopoietic Progenitors Impairs In Vitro Proplatelet Formation and Platelet Release via a Bystander Effect Involving Type I IFN Signaling

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1000847

    JUNV replication in CD34 + cells. CD34 + cells were inoculated with UV-irradiated JUNV or JUNV and stimulated with TPO for ten days. Viral replication was assayed by RT-PCR, immunofluorescence and flow cytometry. (A) RT-PCR studies. (B) To detect JUNV antigens by immunofluorescence cells were washed, cytocentrifuged on silanized glasses, fixed, permeabilized and incubated first with a pool of specific mAbs against JUNV and a rabbit-anti-human vWF polyclonal Ab to identify megakaryocytes, and then with FITC-conjugated anti-rabbit (green) and Cy3-conjugated anti-mouse Igs (red). The slides were counterstained with DAPI and photographed at 1000× magnification. (C) Cells were stained as in B and then analyzed by flow cytometry. (D) As a positive control, JUNV-susceptible Vero-76 cells were inoculated with JUNV and seven days later were stained with the pool of specific mAbs against JUNV followed by Cy3-anti-mouse Igs. Negative controls in B and C were performed by incubating cells only with secondary Abs. Panels show a representative experiment of three similar replicates.
    Figure Legend Snippet: JUNV replication in CD34 + cells. CD34 + cells were inoculated with UV-irradiated JUNV or JUNV and stimulated with TPO for ten days. Viral replication was assayed by RT-PCR, immunofluorescence and flow cytometry. (A) RT-PCR studies. (B) To detect JUNV antigens by immunofluorescence cells were washed, cytocentrifuged on silanized glasses, fixed, permeabilized and incubated first with a pool of specific mAbs against JUNV and a rabbit-anti-human vWF polyclonal Ab to identify megakaryocytes, and then with FITC-conjugated anti-rabbit (green) and Cy3-conjugated anti-mouse Igs (red). The slides were counterstained with DAPI and photographed at 1000× magnification. (C) Cells were stained as in B and then analyzed by flow cytometry. (D) As a positive control, JUNV-susceptible Vero-76 cells were inoculated with JUNV and seven days later were stained with the pool of specific mAbs against JUNV followed by Cy3-anti-mouse Igs. Negative controls in B and C were performed by incubating cells only with secondary Abs. Panels show a representative experiment of three similar replicates.

    Techniques Used: Irradiation, Reverse Transcription Polymerase Chain Reaction, Immunofluorescence, Flow Cytometry, Cytometry, Incubation, Staining, Positive Control

    4) Product Images from "Stability of Porcine Epidemic Diarrhea Virus on Fomite Materials at Different Temperatures"

    Article Title: Stability of Porcine Epidemic Diarrhea Virus on Fomite Materials at Different Temperatures

    Journal: Veterinary Sciences

    doi: 10.3390/vetsci5010021

    Light microscopic images showing distribution and characteristics of PEDV plaques in immunoplaque assay. ( A ) Virus-positive well showing plaques on Vero 76 cells infected with PEDV; ( B ) un-infected control well; ( C ) high magnification of a typical plaque showing viral-induced syncytium (Scale bar: 100 μm); ( D ) comparison of immunoplaque assay with and without agarose overlay. Data presented are average ± SEM from two independent experiments.
    Figure Legend Snippet: Light microscopic images showing distribution and characteristics of PEDV plaques in immunoplaque assay. ( A ) Virus-positive well showing plaques on Vero 76 cells infected with PEDV; ( B ) un-infected control well; ( C ) high magnification of a typical plaque showing viral-induced syncytium (Scale bar: 100 μm); ( D ) comparison of immunoplaque assay with and without agarose overlay. Data presented are average ± SEM from two independent experiments.

    Techniques Used: Infection

    5) Product Images from "Stability of Porcine Epidemic Diarrhea Virus on Fomite Materials at Different Temperatures"

    Article Title: Stability of Porcine Epidemic Diarrhea Virus on Fomite Materials at Different Temperatures

    Journal: Veterinary Sciences

    doi: 10.3390/vetsci5010021

    Light microscopic images showing distribution and characteristics of PEDV plaques in immunoplaque assay. ( A ) Virus-positive well showing plaques on Vero 76 cells infected with PEDV; ( B ) un-infected control well; ( C ) high magnification of a typical plaque showing viral-induced syncytium (Scale bar: 100 μm); ( D ) comparison of immunoplaque assay with and without agarose overlay. Data presented are average ± SEM from two independent experiments.
    Figure Legend Snippet: Light microscopic images showing distribution and characteristics of PEDV plaques in immunoplaque assay. ( A ) Virus-positive well showing plaques on Vero 76 cells infected with PEDV; ( B ) un-infected control well; ( C ) high magnification of a typical plaque showing viral-induced syncytium (Scale bar: 100 μm); ( D ) comparison of immunoplaque assay with and without agarose overlay. Data presented are average ± SEM from two independent experiments.

    Techniques Used: Infection

    6) Product Images from "Stability of Porcine Epidemic Diarrhea Virus on Fomite Materials at Different Temperatures"

    Article Title: Stability of Porcine Epidemic Diarrhea Virus on Fomite Materials at Different Temperatures

    Journal: Veterinary Sciences

    doi: 10.3390/vetsci5010021

    Light microscopic images showing distribution and characteristics of PEDV plaques in immunoplaque assay. ( A ) Virus-positive well showing plaques on Vero 76 cells infected with PEDV; ( B ) un-infected control well; ( C ) high magnification of a typical plaque showing viral-induced syncytium (Scale bar: 100 μm); ( D ) comparison of immunoplaque assay with and without agarose overlay. Data presented are average ± SEM from two independent experiments.
    Figure Legend Snippet: Light microscopic images showing distribution and characteristics of PEDV plaques in immunoplaque assay. ( A ) Virus-positive well showing plaques on Vero 76 cells infected with PEDV; ( B ) un-infected control well; ( C ) high magnification of a typical plaque showing viral-induced syncytium (Scale bar: 100 μm); ( D ) comparison of immunoplaque assay with and without agarose overlay. Data presented are average ± SEM from two independent experiments.

    Techniques Used: Infection

    7) Product Images from "Discovery of a Novel Compound with Anti-Venezuelan Equine Encephalitis Virus Activity That Targets the Nonstructural Protein 2"

    Article Title: Discovery of a Novel Compound with Anti-Venezuelan Equine Encephalitis Virus Activity That Targets the Nonstructural Protein 2

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1004213

    Anti-VEEV activity of CID15997213. ( A ) Dose response curve of CID15997213 in the CPE-based anti-VEEV TC-83 assay using Vero 76 cells from a representative experiment. ( B ) Titer reduction assay results for CID15997213. Vero 76 cells grown in 6-well plates were infected with 0.05 MOI of TC-83 or TrD and then incubated in the presence of CID15997213 at the denoted concentrations. Forty hours later the supernatant was harvested and the titer of the progeny virus was determined. Each point represents the mean from two independent experiments. The horizontal line in the figure indicates the detection limit of the assay. ( C , D ) Viral RNA and protein analysis. Vero 76 cells were infected with VEEV TC-83 at MOI of 5 and then incubated in the presence of DMSO or CID15997213 for 16 hours. In C , viral RNA was quantified using a quantitative real-time RT-PCR method with the total RNA from the cells. In D , proteins from VEEV TC83-infected cell extracts were analyzed by western blot assay. Closed triangles indicate bands corresponding to actin (internal control) and open arrows indicate bands that are specific to the viral proteins.
    Figure Legend Snippet: Anti-VEEV activity of CID15997213. ( A ) Dose response curve of CID15997213 in the CPE-based anti-VEEV TC-83 assay using Vero 76 cells from a representative experiment. ( B ) Titer reduction assay results for CID15997213. Vero 76 cells grown in 6-well plates were infected with 0.05 MOI of TC-83 or TrD and then incubated in the presence of CID15997213 at the denoted concentrations. Forty hours later the supernatant was harvested and the titer of the progeny virus was determined. Each point represents the mean from two independent experiments. The horizontal line in the figure indicates the detection limit of the assay. ( C , D ) Viral RNA and protein analysis. Vero 76 cells were infected with VEEV TC-83 at MOI of 5 and then incubated in the presence of DMSO or CID15997213 for 16 hours. In C , viral RNA was quantified using a quantitative real-time RT-PCR method with the total RNA from the cells. In D , proteins from VEEV TC83-infected cell extracts were analyzed by western blot assay. Closed triangles indicate bands corresponding to actin (internal control) and open arrows indicate bands that are specific to the viral proteins.

    Techniques Used: Activity Assay, Infection, Incubation, Quantitative RT-PCR, Western Blot

    8) Product Images from "Inhibition of severe acute respiratory syndrome coronavirus replication in a lethal SARS-CoV BALB/c mouse model by stinging nettle lectin, Urtica dioica agglutinin"

    Article Title: Inhibition of severe acute respiratory syndrome coronavirus replication in a lethal SARS-CoV BALB/c mouse model by stinging nettle lectin, Urtica dioica agglutinin

    Journal: Antiviral research

    doi: 10.1016/j.antiviral.2011.02.003

    A . Effect of live SARS-CoV infection by UDA (virus titers: log 10 CCID 50 per 0.18 ml). SARS-CoV was exposed to UDA for 1 h and the UDA-exposed virus suspension was titrated in Vero 76 cells.
    Figure Legend Snippet: A . Effect of live SARS-CoV infection by UDA (virus titers: log 10 CCID 50 per 0.18 ml). SARS-CoV was exposed to UDA for 1 h and the UDA-exposed virus suspension was titrated in Vero 76 cells.

    Techniques Used: Infection

    9) Product Images from "Stability of Porcine Epidemic Diarrhea Virus on Fomite Materials at Different Temperatures"

    Article Title: Stability of Porcine Epidemic Diarrhea Virus on Fomite Materials at Different Temperatures

    Journal: Veterinary Sciences

    doi: 10.3390/vetsci5010021

    Light microscopic images showing distribution and characteristics of PEDV plaques in immunoplaque assay. ( A ) Virus-positive well showing plaques on Vero 76 cells infected with PEDV; ( B ) un-infected control well; ( C ) high magnification of a typical plaque showing viral-induced syncytium (Scale bar: 100 μm); ( D ) comparison of immunoplaque assay with and without agarose overlay. Data presented are average ± SEM from two independent experiments.
    Figure Legend Snippet: Light microscopic images showing distribution and characteristics of PEDV plaques in immunoplaque assay. ( A ) Virus-positive well showing plaques on Vero 76 cells infected with PEDV; ( B ) un-infected control well; ( C ) high magnification of a typical plaque showing viral-induced syncytium (Scale bar: 100 μm); ( D ) comparison of immunoplaque assay with and without agarose overlay. Data presented are average ± SEM from two independent experiments.

    Techniques Used: Infection

    Related Articles

    Plaque Assay:

    Article Title: Gene expression profiling of rubella virus infected primary endothelial cells of fetal and adult origin
    Article Snippet: .. Virus titer of the supernatant was determined by immunocolorimetric plaque assay on Vero76 cells in duplicate. .. Microarray, hybridization and gene expression analysis For microarray analysis, 2°106 endothelial cells of fetal and adult origin were infected with RV at an MOI of 10 and cells were lysed 36 h post infection with 600 μl RLT buffer from the RNAeasy kit (Qiagen).

    Neutralization:

    Article Title: Evaluation of biological safety in vitro and immunogenicity in vivo of recombinant Escherichia coli Shiga toxoids as candidate vaccines in cattle
    Article Snippet: .. Vero cell cytotoxicity assay (VCA) and Vero cell cytotoxicity neutralization assay (VNA) The VCA was performed in 96-well microtiter plates (Nunc, Wiesbaden, Germany) using Vero cells (ATCC CRL 1587, LGC-Promochem GmbH, Wesel, Germany) as previously described [ ] to determine the cytotoxicity (verocytotoxic doses 50%, CD50 /mL) of the rStxWT preparations and for adjustment of stock solutions (20 000 CD50 /mL). .. The VNA was used for the determination of the neutralization activity in serum of vaccinated calves and was done as previously described [ ] in order to determine the titre of neutralizing antibodies [nAb titre] against either wild type Stx1 or wild type Stx2 (Sigma-Aldrich Chemie GmbH, Taufkirchen, Germany).

    Infection:

    Article Title: Jun?n Virus Infection of Human Hematopoietic Progenitors Impairs In Vitro Proplatelet Formation and Platelet Release via a Bystander Effect Involving Type I IFN Signaling
    Article Snippet: .. Cell infection CD34+ cells (1×104 ) or a monolayer of Vero-76 cells were inoculated with JUNV at a multiplicity of infection (MOI) of one or the equivalent volume of UV-irradiated virus for 1 hr at 37°C. ..

    Cytotoxicity Assay:

    Article Title: Evaluation of biological safety in vitro and immunogenicity in vivo of recombinant Escherichia coli Shiga toxoids as candidate vaccines in cattle
    Article Snippet: .. Vero cell cytotoxicity assay (VCA) and Vero cell cytotoxicity neutralization assay (VNA) The VCA was performed in 96-well microtiter plates (Nunc, Wiesbaden, Germany) using Vero cells (ATCC CRL 1587, LGC-Promochem GmbH, Wesel, Germany) as previously described [ ] to determine the cytotoxicity (verocytotoxic doses 50%, CD50 /mL) of the rStxWT preparations and for adjustment of stock solutions (20 000 CD50 /mL). .. The VNA was used for the determination of the neutralization activity in serum of vaccinated calves and was done as previously described [ ] in order to determine the titre of neutralizing antibodies [nAb titre] against either wild type Stx1 or wild type Stx2 (Sigma-Aldrich Chemie GmbH, Taufkirchen, Germany).

    other:

    Article Title: Jun?n Virus Infection of Human Hematopoietic Progenitors Impairs In Vitro Proplatelet Formation and Platelet Release via a Bystander Effect Involving Type I IFN Signaling
    Article Snippet: Vero-76 cells Monolayers of Vero-76 cells (CRL-1587, the American Type Culture Collection (ATCC), Manassas, VA) were grown in minimum essential medium (MEM) containing 10% fetal calf serum (FCS) and antibiotics.

    Activity Assay:

    Article Title: The antiviral activity of naturally occurring proteins and their peptide fragments after chemical modification
    Article Snippet: .. Vero76 cells, used to measure the antiviral activity against HSV-1, Madin–Darby bovine kidney cells (MDBK) and Swine Testicular (ST) cells, used to measure the inhibitory effect of the chemically modified proteins against PIV-3 and PRCV, respectively, were originally purchased from ATCC. ..

    Modification:

    Article Title: Risk1, a Phosphatidylinositol 3-Kinase Effector, Promotes Rickettsia typhi Intracellular Survival
    Article Snippet: .. Vero 76 (African green monkey kidney, RL-1587; ATCC) and HeLa (CCL-2; ATCC) cells were maintained in minimal Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS) at 37°C with 5% CO2 . ..

    Article Title: The antiviral activity of naturally occurring proteins and their peptide fragments after chemical modification
    Article Snippet: .. Vero76 cells, used to measure the antiviral activity against HSV-1, Madin–Darby bovine kidney cells (MDBK) and Swine Testicular (ST) cells, used to measure the inhibitory effect of the chemically modified proteins against PIV-3 and PRCV, respectively, were originally purchased from ATCC. ..

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  • 99
    ATCC vero e6 cells
    Radio-uptake of 99m Tc-pertechnetate by planar scintigraphy. (a) Experimental overview of in vitro evaluation of the rMERS-CoV/ hNIS virus. <t>Vero</t> <t>E6</t> cells were infected with rMERS-CoV or rMERS-CoV/ hNIS at an MOI of 0.01 or 0.04. At various time points postinfection, the cells were incubated with 99m Tc-pertechnetate, and images of the plates were acquired. (b) Plate layout for hNIS functional assays. (c) Representative images of the plates acquired at 24 h postinfection at an MOI of 0.01 (top plates) or 0.04 (bottom plates) after incubation with 99m Tc-pertechnetate.
    Vero E6 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 113 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/vero e6 cells/product/ATCC
    Average 99 stars, based on 113 article reviews
    Price from $9.99 to $1999.99
    vero e6 cells - by Bioz Stars, 2020-11
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    Radio-uptake of 99m Tc-pertechnetate by planar scintigraphy. (a) Experimental overview of in vitro evaluation of the rMERS-CoV/ hNIS virus. Vero E6 cells were infected with rMERS-CoV or rMERS-CoV/ hNIS at an MOI of 0.01 or 0.04. At various time points postinfection, the cells were incubated with 99m Tc-pertechnetate, and images of the plates were acquired. (b) Plate layout for hNIS functional assays. (c) Representative images of the plates acquired at 24 h postinfection at an MOI of 0.01 (top plates) or 0.04 (bottom plates) after incubation with 99m Tc-pertechnetate.

    Journal: mSphere

    Article Title: The Human Sodium Iodide Symporter as a Reporter Gene for Studying Middle East Respiratory Syndrome Coronavirus Pathogenesis

    doi: 10.1128/mSphere.00540-18

    Figure Lengend Snippet: Radio-uptake of 99m Tc-pertechnetate by planar scintigraphy. (a) Experimental overview of in vitro evaluation of the rMERS-CoV/ hNIS virus. Vero E6 cells were infected with rMERS-CoV or rMERS-CoV/ hNIS at an MOI of 0.01 or 0.04. At various time points postinfection, the cells were incubated with 99m Tc-pertechnetate, and images of the plates were acquired. (b) Plate layout for hNIS functional assays. (c) Representative images of the plates acquired at 24 h postinfection at an MOI of 0.01 (top plates) or 0.04 (bottom plates) after incubation with 99m Tc-pertechnetate.

    Article Snippet: Vero E6 cells (ATCC CRL-1586) were maintained in Dulbecco’s modified Eagle medium (DMEM) (Lonza) supplemented with 5% fetal bovine serum (FBS) and incubated at 37°C and 5% CO2 without antibiotics or antimycotics.

    Techniques: In Vitro, Infection, Incubation, Functional Assay

    Retention of hNIS transgene following viral kinetics analysis and serial passage. (a and b) Vero E6 cells were infected with rMERS-CoV/hNIS (a) or parental rMERS-CoV (b) at an MOI of 0.01 or 3 and then collected at 96 h postinfection for RT-PCR. (c) Retention of the hNIS gene following serial passage. RNA was extracted from cells 72 h postinfection followed by RT-PCR at passage 6. A positive-control virus (C+) and uninfected negative-control cells (C−) were used as controls.

    Journal: mSphere

    Article Title: The Human Sodium Iodide Symporter as a Reporter Gene for Studying Middle East Respiratory Syndrome Coronavirus Pathogenesis

    doi: 10.1128/mSphere.00540-18

    Figure Lengend Snippet: Retention of hNIS transgene following viral kinetics analysis and serial passage. (a and b) Vero E6 cells were infected with rMERS-CoV/hNIS (a) or parental rMERS-CoV (b) at an MOI of 0.01 or 3 and then collected at 96 h postinfection for RT-PCR. (c) Retention of the hNIS gene following serial passage. RNA was extracted from cells 72 h postinfection followed by RT-PCR at passage 6. A positive-control virus (C+) and uninfected negative-control cells (C−) were used as controls.

    Article Snippet: Vero E6 cells (ATCC CRL-1586) were maintained in Dulbecco’s modified Eagle medium (DMEM) (Lonza) supplemented with 5% fetal bovine serum (FBS) and incubated at 37°C and 5% CO2 without antibiotics or antimycotics.

    Techniques: Infection, Reverse Transcription Polymerase Chain Reaction, Positive Control, Negative Control

    Kinetics of rMERS-CoV/ hNIS and parental rMERS-CoV replication in Vero E6 cells. (a and b) Multistep (a) and one-step (b) growth curves of Vero E6 cells infected with rMERS-CoV (Parental) and rMERS-CoV/ hNIS ( hNIS ). Quantification of the release of infectious virus at the indicated time points (hours postexposure) was determined by plaque assays. Each data point represents the mean ± standard deviation (SD) (error bar) averaged from three independent experiments. (c and d) Cytopathology of rMERS-CoV and rMERS-CoV/ hNIS in Vero E6 cells. The cells were infected with either rMERS-CoV or rMERS-CoV/ hNIS at an MOI of 0.01 (c) or 3 (d) and analyzed by light microscopy at the indicated time points. Photomicrographs were taken using a 40× objective.

    Journal: mSphere

    Article Title: The Human Sodium Iodide Symporter as a Reporter Gene for Studying Middle East Respiratory Syndrome Coronavirus Pathogenesis

    doi: 10.1128/mSphere.00540-18

    Figure Lengend Snippet: Kinetics of rMERS-CoV/ hNIS and parental rMERS-CoV replication in Vero E6 cells. (a and b) Multistep (a) and one-step (b) growth curves of Vero E6 cells infected with rMERS-CoV (Parental) and rMERS-CoV/ hNIS ( hNIS ). Quantification of the release of infectious virus at the indicated time points (hours postexposure) was determined by plaque assays. Each data point represents the mean ± standard deviation (SD) (error bar) averaged from three independent experiments. (c and d) Cytopathology of rMERS-CoV and rMERS-CoV/ hNIS in Vero E6 cells. The cells were infected with either rMERS-CoV or rMERS-CoV/ hNIS at an MOI of 0.01 (c) or 3 (d) and analyzed by light microscopy at the indicated time points. Photomicrographs were taken using a 40× objective.

    Article Snippet: Vero E6 cells (ATCC CRL-1586) were maintained in Dulbecco’s modified Eagle medium (DMEM) (Lonza) supplemented with 5% fetal bovine serum (FBS) and incubated at 37°C and 5% CO2 without antibiotics or antimycotics.

    Techniques: Infection, Standard Deviation, Light Microscopy

    Stability of the GFP reporter gene expressed by rLASV-GFP. Cells were exposed to rLASV-GFP (MOI = 0.01). At 72 h PI, TCS were collected (passage 1, P1), and virus titers were determined by plaque assay. Fresh Vero ( a ) or Vero-E6 cells ( b ) were exposed to with TCS from P1 (MOI = 0.01). This process was serially repeated throughout P10. GFP expression (green) was measured by high-content imaging. Nuclei were stained with DAPI (blue). Viral RNA was extracted from TCS of P1 to P10 to amplify DNA fragments containing an GFP-P2A and a part of the NP ORF using RT-PCR. Images are representative field images of individual wells. Bar, 200 µm.

    Journal: Viruses

    Article Title: Recombinant Lassa Virus Expressing Green Fluorescent Protein as a Tool for High-Throughput Drug Screens and Neutralizing Antibody Assays

    doi: 10.3390/v10110655

    Figure Lengend Snippet: Stability of the GFP reporter gene expressed by rLASV-GFP. Cells were exposed to rLASV-GFP (MOI = 0.01). At 72 h PI, TCS were collected (passage 1, P1), and virus titers were determined by plaque assay. Fresh Vero ( a ) or Vero-E6 cells ( b ) were exposed to with TCS from P1 (MOI = 0.01). This process was serially repeated throughout P10. GFP expression (green) was measured by high-content imaging. Nuclei were stained with DAPI (blue). Viral RNA was extracted from TCS of P1 to P10 to amplify DNA fragments containing an GFP-P2A and a part of the NP ORF using RT-PCR. Images are representative field images of individual wells. Bar, 200 µm.

    Article Snippet: Human ( Homo sapiens ) adenocarcinoma alveolar basal epithelial A549 (American Type Culture Collection [ATCC], Manassas, VA; #CCL-185) cells, epithelial Hela cells (ATCC, #CCL-2), and hepatocarcinoma Huh-7 cells (a kind gift from Hideki Ebihara, Rocky Mountain Laboratories, Hamilton, MT, USA), and grivet ( Chlorocebus aethiops ) kidney epithelial Vero cells (ATCC, #CCL-81), and Vero E6 cells (ATCC, #CRL-1586) were grown in Gibco Dulbecco’s modified Eagle’s medium (DMEM) (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS, Sigma-Aldrich, St. Louis, MO, USA).

    Techniques: Plaque Assay, Expressing, Imaging, Staining, Reverse Transcription Polymerase Chain Reaction

    Antiviral drug evaluation based on recombinant LASV expressing GFP (rLASV-GFP). ( a ) Vero E6 cells were pretreated with drugs at the indicated concentrations and then exposed to rLASV-WT or rLASV-GFP (MOI = 0.1) in the presence of the drugs. Viral titers in TCS at 48 h PI were determined by plaque assay. Values represent the means ± standard deviations of triplicate samples. ( b ) Infectivity of rLASV-GFP in A549, HeLa, Huh7, and Vero E6 cells at the indicated MOIs at 24, 48, and 72h PI as determined by GFP-expression using high-content imaging. ( c ) Effect of favipiravir and ribavirin on rLASV-GFP multiplication at 48 (orange filled circles) and 72 h PI (green filled squares). Cells were exposed to rLASV-GFP (MOI = 0.1) and treated with various concentrations of favipiravir or ribavirin. The percentage of GFP-positive cells was determined at 48 h or 72 h PI. ( d ) Half-maximal effective concentrations (EC 50 ) of favipiravir and ribavirin to inhibit rLASV-GFP infection in four cell types at 48 and 72 h PI.

    Journal: Viruses

    Article Title: Recombinant Lassa Virus Expressing Green Fluorescent Protein as a Tool for High-Throughput Drug Screens and Neutralizing Antibody Assays

    doi: 10.3390/v10110655

    Figure Lengend Snippet: Antiviral drug evaluation based on recombinant LASV expressing GFP (rLASV-GFP). ( a ) Vero E6 cells were pretreated with drugs at the indicated concentrations and then exposed to rLASV-WT or rLASV-GFP (MOI = 0.1) in the presence of the drugs. Viral titers in TCS at 48 h PI were determined by plaque assay. Values represent the means ± standard deviations of triplicate samples. ( b ) Infectivity of rLASV-GFP in A549, HeLa, Huh7, and Vero E6 cells at the indicated MOIs at 24, 48, and 72h PI as determined by GFP-expression using high-content imaging. ( c ) Effect of favipiravir and ribavirin on rLASV-GFP multiplication at 48 (orange filled circles) and 72 h PI (green filled squares). Cells were exposed to rLASV-GFP (MOI = 0.1) and treated with various concentrations of favipiravir or ribavirin. The percentage of GFP-positive cells was determined at 48 h or 72 h PI. ( d ) Half-maximal effective concentrations (EC 50 ) of favipiravir and ribavirin to inhibit rLASV-GFP infection in four cell types at 48 and 72 h PI.

    Article Snippet: Human ( Homo sapiens ) adenocarcinoma alveolar basal epithelial A549 (American Type Culture Collection [ATCC], Manassas, VA; #CCL-185) cells, epithelial Hela cells (ATCC, #CCL-2), and hepatocarcinoma Huh-7 cells (a kind gift from Hideki Ebihara, Rocky Mountain Laboratories, Hamilton, MT, USA), and grivet ( Chlorocebus aethiops ) kidney epithelial Vero cells (ATCC, #CCL-81), and Vero E6 cells (ATCC, #CRL-1586) were grown in Gibco Dulbecco’s modified Eagle’s medium (DMEM) (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS, Sigma-Aldrich, St. Louis, MO, USA).

    Techniques: Recombinant, Expressing, Plaque Assay, Infection, Imaging

    Single-step growth curves of the NSP6-deficient viruses in IFN-α/β-deficient Vero E6 cells. Vero E6 cells were infected with rSA11 or rSA11-delNSP6 (A) or with rSA11-s11KU or rSA11-s11KU-delNSP6 (B) at an MOI of 5 and then incubated for various times. The virus titers in the cultures were determined by plaque assay. The data shown are the mean viral titers and SDs from three independent cell cultures.

    Journal: Journal of Virology

    Article Title: Reverse Genetics System Demonstrates that Rotavirus Nonstructural Protein NSP6 Is Not Essential for Viral Replication in Cell Culture

    doi: 10.1128/JVI.00695-17

    Figure Lengend Snippet: Single-step growth curves of the NSP6-deficient viruses in IFN-α/β-deficient Vero E6 cells. Vero E6 cells were infected with rSA11 or rSA11-delNSP6 (A) or with rSA11-s11KU or rSA11-s11KU-delNSP6 (B) at an MOI of 5 and then incubated for various times. The virus titers in the cultures were determined by plaque assay. The data shown are the mean viral titers and SDs from three independent cell cultures.

    Article Snippet: Monkey kidney cell lines, MA104, CV-1, and Vero E6 (CRL-1586; American Type Culture Collection), were cultured in complete medium.

    Techniques: Infection, Incubation, Plaque Assay

    (A) Indirect immunofluorescence detection of MRV in Vero E6 cells infected with strain B/03. The cells were fixed with 4% paraformaldehyde, blocked with 1% BSA, washed, and incubated with mouse anti-MRV (T3D) antibody. The cells were then incubated with FITC-labeled goat anti-mouse IgG secondary antibody. (B) Electron micrographs. a: Negative staining of cell culture supernatant. Non-enveloped reoviral-like particles with double-layered capsid structure were observed (diameter = approximately 70 nm). b: Ultra-thin sections of infected Vero E6 cells displayed typical contrast-rich virus particles, organized as paracrystalline structures within the cytosol. (C) SDS-PAGE demonstrating electrophoresis pattern. L, M and S represent large, medium and small segments, respectively. dsRNA segments were separated by electrophoresis in 8% ( w / v ) polyacrylamide slab gels.

    Journal: Infection, Genetics and Evolution

    Article Title: Characterization and pathogenicity of a novel mammalian orthoreovirus from wild short-nosed fruit bats

    doi: 10.1016/j.meegid.2016.05.039

    Figure Lengend Snippet: (A) Indirect immunofluorescence detection of MRV in Vero E6 cells infected with strain B/03. The cells were fixed with 4% paraformaldehyde, blocked with 1% BSA, washed, and incubated with mouse anti-MRV (T3D) antibody. The cells were then incubated with FITC-labeled goat anti-mouse IgG secondary antibody. (B) Electron micrographs. a: Negative staining of cell culture supernatant. Non-enveloped reoviral-like particles with double-layered capsid structure were observed (diameter = approximately 70 nm). b: Ultra-thin sections of infected Vero E6 cells displayed typical contrast-rich virus particles, organized as paracrystalline structures within the cytosol. (C) SDS-PAGE demonstrating electrophoresis pattern. L, M and S represent large, medium and small segments, respectively. dsRNA segments were separated by electrophoresis in 8% ( w / v ) polyacrylamide slab gels.

    Article Snippet: Vero E6 cells were obtained from the ATCC (ATCC® CRL-1586™) and grown at 37 °C in 5% CO2 in DMEM supplemented with 2 mM glutamine, 5% fetal calf serum and antibiotics.

    Techniques: Immunofluorescence, Infection, Incubation, Labeling, Negative Staining, Cell Culture, SDS Page, Electrophoresis