vero cells  (Millipore)

 
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    Name:
    Vero cells
    Description:
    This product is provided as delivered and specified by the issuing Pharmacopoeia All information provided in support of this product including SDS and any product information leaflets have been developed and issued under the Authority of the issuing Pharmacopoeia For further information and support please go to the website of the issuing Pharmacopoeia
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    Structured Review

    Millipore vero cells
    FACS analyses on <t>A549,</t> fibroblasts, and <t>Vero</t> cells for the expression of CD55, CD59, and CD46. Detection of membrane-anchored complement regulator protein (mCRP) expression on ZIKV-producing cells by FACS analysis. Cells were stained with anti-human CD46, CD55, or CD59 (dark grey histogram) or isotype-matched Ab (light gray histogram).
    This product is provided as delivered and specified by the issuing Pharmacopoeia All information provided in support of this product including SDS and any product information leaflets have been developed and issued under the Authority of the issuing Pharmacopoeia For further information and support please go to the website of the issuing Pharmacopoeia
    https://www.bioz.com/result/vero cells/product/Millipore
    Average 99 stars, based on 1 article reviews
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    vero cells - by Bioz Stars, 2021-06
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    Images

    1) Product Images from "Incorporation of CD55 into the Zika Viral Envelope Contributes to Its Stability against Human Complement"

    Article Title: Incorporation of CD55 into the Zika Viral Envelope Contributes to Its Stability against Human Complement

    Journal: Viruses

    doi: 10.3390/v13030510

    FACS analyses on A549, fibroblasts, and Vero cells for the expression of CD55, CD59, and CD46. Detection of membrane-anchored complement regulator protein (mCRP) expression on ZIKV-producing cells by FACS analysis. Cells were stained with anti-human CD46, CD55, or CD59 (dark grey histogram) or isotype-matched Ab (light gray histogram).
    Figure Legend Snippet: FACS analyses on A549, fibroblasts, and Vero cells for the expression of CD55, CD59, and CD46. Detection of membrane-anchored complement regulator protein (mCRP) expression on ZIKV-producing cells by FACS analysis. Cells were stained with anti-human CD46, CD55, or CD59 (dark grey histogram) or isotype-matched Ab (light gray histogram).

    Techniques Used: FACS, Expressing, Staining

    Serum stability of Zika virus (ZIKV) is affected by cell-line-dependent features. ( A ) ZIKV Insect , ZIKV A549 , ZIKV Fibro, and ZIKV Vero were incubated with increasing amounts of active or heat-inactivated human serum. ( B ) For analysis of the different viral preparations, the input virus was normalized on the amount of virus in heat-inactivated normal human serum (hiNHS). The graph shows the stability of ZIKV derived from different cells in 50%NHS. In both sets of experiments, virus-serum mixtures were incubated for 1 h at 37 °C, then diluted 10-fold, titrated on 12-well plates of overnight-plated Vero cells and incubated for 1 h before plaque agarose was overlaid. Plaques were visualized four days post infection using crystal violet staining. All assays were performed in triplicate, and the error bars show standard deviations. A 95% significance level ( p
    Figure Legend Snippet: Serum stability of Zika virus (ZIKV) is affected by cell-line-dependent features. ( A ) ZIKV Insect , ZIKV A549 , ZIKV Fibro, and ZIKV Vero were incubated with increasing amounts of active or heat-inactivated human serum. ( B ) For analysis of the different viral preparations, the input virus was normalized on the amount of virus in heat-inactivated normal human serum (hiNHS). The graph shows the stability of ZIKV derived from different cells in 50%NHS. In both sets of experiments, virus-serum mixtures were incubated for 1 h at 37 °C, then diluted 10-fold, titrated on 12-well plates of overnight-plated Vero cells and incubated for 1 h before plaque agarose was overlaid. Plaques were visualized four days post infection using crystal violet staining. All assays were performed in triplicate, and the error bars show standard deviations. A 95% significance level ( p

    Techniques Used: Incubation, Derivative Assay, Infection, Staining

    Western blot of ZIKV lysates confirms CD55 uptake. ZIKV-containing supernatants (concentrated) of infected A549, Vero, or Insect cells were characterized for CD55 incorporation by Western blotting. Samples were incubated with anti-human CD55 antibody. As load control, anti-ZIKV envelope was used. A representative Western blot analysis is shown.
    Figure Legend Snippet: Western blot of ZIKV lysates confirms CD55 uptake. ZIKV-containing supernatants (concentrated) of infected A549, Vero, or Insect cells were characterized for CD55 incorporation by Western blotting. Samples were incubated with anti-human CD55 antibody. As load control, anti-ZIKV envelope was used. A representative Western blot analysis is shown.

    Techniques Used: Western Blot, Infection, Incubation

    Capture ELISA indicates CD55 incorporation in ZIKV envelope. Graphs depict fold changes of RNA genome copies of captured ZIKV, analyzed by RT-PCR. Intact virions ZIKV A549 , ZIKV Insect , ZIKV Fibro , or ZIKV Vero were incubated overnight on pre-coated ELISA plates, bearing antibodies against human CD46, CD55, and CD59. ZIKV envelope served as a positive control. Isotype control was used as a negative control. After extensive washing steps, in order to remove unbound virions, ZIKV particles were lysed and RNA was extracted. The captured copy number of ZIKV genomes was characterized by determining the threshold-cycles by qPCR. Experiments were done two times in duplicates, and the error bars show standard deviations. Data were analyzed by ANOVA followed by Sidak’s multiple comparisons. A 95% significance level ( p
    Figure Legend Snippet: Capture ELISA indicates CD55 incorporation in ZIKV envelope. Graphs depict fold changes of RNA genome copies of captured ZIKV, analyzed by RT-PCR. Intact virions ZIKV A549 , ZIKV Insect , ZIKV Fibro , or ZIKV Vero were incubated overnight on pre-coated ELISA plates, bearing antibodies against human CD46, CD55, and CD59. ZIKV envelope served as a positive control. Isotype control was used as a negative control. After extensive washing steps, in order to remove unbound virions, ZIKV particles were lysed and RNA was extracted. The captured copy number of ZIKV genomes was characterized by determining the threshold-cycles by qPCR. Experiments were done two times in duplicates, and the error bars show standard deviations. Data were analyzed by ANOVA followed by Sidak’s multiple comparisons. A 95% significance level ( p

    Techniques Used: Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Incubation, Positive Control, Negative Control, Real-time Polymerase Chain Reaction

    Effect of IgM on complement activation. To investigate whether natural serum IgMs affect the complement activation of human cell-derived ZIKV, anti-human IgM blocking antibodies were incubated in 50% NHS or hiNHS for 30 min on ice, before ZIKV A549 or ZIKV Fibro were added. The experimental conditions are indicated by “x”. As positive control, ZIKV insect was included [ 13 ]. After incubation for 1 h at 37 °C, the virus–serum mixture was serial diluted and titrated on 12-well plates of overnight-plated Vero cells. One hour after incubation at 37 °C, plaque agarose was overlaid. Viral concentration was determined and calculated four days post-infection using crystal violet staining. All virus lysis experiments were conducted in triplicate, and the error bars show standard deviations. A 95% significance level ( p
    Figure Legend Snippet: Effect of IgM on complement activation. To investigate whether natural serum IgMs affect the complement activation of human cell-derived ZIKV, anti-human IgM blocking antibodies were incubated in 50% NHS or hiNHS for 30 min on ice, before ZIKV A549 or ZIKV Fibro were added. The experimental conditions are indicated by “x”. As positive control, ZIKV insect was included [ 13 ]. After incubation for 1 h at 37 °C, the virus–serum mixture was serial diluted and titrated on 12-well plates of overnight-plated Vero cells. One hour after incubation at 37 °C, plaque agarose was overlaid. Viral concentration was determined and calculated four days post-infection using crystal violet staining. All virus lysis experiments were conducted in triplicate, and the error bars show standard deviations. A 95% significance level ( p

    Techniques Used: Activation Assay, Derivative Assay, Blocking Assay, Incubation, Positive Control, Concentration Assay, Infection, Staining, Lysis

    2) Product Images from "Specific Association of Glycoprotein B with Lipid Rafts during Herpes Simplex Virus Entry"

    Article Title: Specific Association of Glycoprotein B with Lipid Rafts during Herpes Simplex Virus Entry

    Journal: Journal of Virology

    doi: 10.1128/JVI.77.17.9542-9552.2003

    Effect of cholesterol depletion from cells on the course of HSV entry. Vero cells were infected with HSV-1 (KOS/tk12). At various times following infection (indicated on the abscissa), cells were treated for 30 min with MβCD. The drug was washed out, and medium or cholesterol (chol) was added. After 30 min, the cholesterol was removed, and infection was allowed to proceed. β-Galactosidase activity at 6 h postinfection was determined. The results shown are representative of three independent experiments, and 100% entry represents the β-galactosidase activity at 6 h in the absence of MβCD.
    Figure Legend Snippet: Effect of cholesterol depletion from cells on the course of HSV entry. Vero cells were infected with HSV-1 (KOS/tk12). At various times following infection (indicated on the abscissa), cells were treated for 30 min with MβCD. The drug was washed out, and medium or cholesterol (chol) was added. After 30 min, the cholesterol was removed, and infection was allowed to proceed. β-Galactosidase activity at 6 h postinfection was determined. The results shown are representative of three independent experiments, and 100% entry represents the β-galactosidase activity at 6 h in the absence of MβCD.

    Techniques Used: Infection, Activity Assay

    Effect of cholesterol depletion or chelation on HSV entry. Vero cells (A and B) and B78 A10 and C10 cells (C) were treated with increasing concentrations of MβCD (A and C) or nystatin (B). The drugs were washed out, and the cells were infected with HSV-1 (KOS/tk12) and assayed for β-galactosidase activity at 6 h postinfection. Results presented in A and B are the mean of three independent experiments done in duplicate, with standard deviations. The results shown in panel C are representative of three experiments. A value of 100% entry represents the β-galactosidase activity at 6 h in the absence of MβCD.
    Figure Legend Snippet: Effect of cholesterol depletion or chelation on HSV entry. Vero cells (A and B) and B78 A10 and C10 cells (C) were treated with increasing concentrations of MβCD (A and C) or nystatin (B). The drugs were washed out, and the cells were infected with HSV-1 (KOS/tk12) and assayed for β-galactosidase activity at 6 h postinfection. Results presented in A and B are the mean of three independent experiments done in duplicate, with standard deviations. The results shown in panel C are representative of three experiments. A value of 100% entry represents the β-galactosidase activity at 6 h in the absence of MβCD.

    Techniques Used: Infection, Activity Assay

    Effect of cholesterol depletion from cells on expression of nectin-1 and on attachment and replication of HSV. (A) C10 cells were untreated (−) or treated (+) with 30 mM MβCD for 30 min. The drug was washed out, and HSV-1 (KOS) was allowed to bind to the cells for 1 h at 4°C. Total cell proteins were extracted, resolved by SDS-PAGE, and probed with anti-nectin-1 MAb CK6 (upper panel) or anti-gD MAb DL6 (lower panel). (B) C10 cells were treated with increasing concentrations of MβCD (indicated at the right), then stained with anti-nectin-1 MAb CK41 directly labeled with phycoerythrin, and analyzed by FACS. The control (open area) represents the fluorescence of unstained and drug untreated cells. (C) Vero cells were treated with increasing concentrations of MβCD. The drug was washed out, and the cells were infected with HSV1 (KOS) or vesicular stomatitis virus (VSV) for 24 h at 37°C. Then the cells were fixed, and plaques werevisualized by immunoperoxidase staining (HSV) or crystal violet staining (vesicular stomatitis virus).
    Figure Legend Snippet: Effect of cholesterol depletion from cells on expression of nectin-1 and on attachment and replication of HSV. (A) C10 cells were untreated (−) or treated (+) with 30 mM MβCD for 30 min. The drug was washed out, and HSV-1 (KOS) was allowed to bind to the cells for 1 h at 4°C. Total cell proteins were extracted, resolved by SDS-PAGE, and probed with anti-nectin-1 MAb CK6 (upper panel) or anti-gD MAb DL6 (lower panel). (B) C10 cells were treated with increasing concentrations of MβCD (indicated at the right), then stained with anti-nectin-1 MAb CK41 directly labeled with phycoerythrin, and analyzed by FACS. The control (open area) represents the fluorescence of unstained and drug untreated cells. (C) Vero cells were treated with increasing concentrations of MβCD. The drug was washed out, and the cells were infected with HSV1 (KOS) or vesicular stomatitis virus (VSV) for 24 h at 37°C. Then the cells were fixed, and plaques werevisualized by immunoperoxidase staining (HSV) or crystal violet staining (vesicular stomatitis virus).

    Techniques Used: Expressing, SDS Page, Staining, Labeling, FACS, Fluorescence, Infection, Immunoperoxidase Staining

    Effect of cholesterol replenishment on HSV entry. Vero cells were untreated (left, rectangles) or treated (right, rectangles) with 7.5 mM MβCD for 30 min. MβCD was washed out, and various amounts of cholesterol in MβCD were added. After 30 min, the cholesterol was removed, and the cells were infected with HSV-1 (KOS/tk12) and assayed for β-galactosidase activity at 6 h postinfection. The results presented are representative of two independent experiments done in triplicate.
    Figure Legend Snippet: Effect of cholesterol replenishment on HSV entry. Vero cells were untreated (left, rectangles) or treated (right, rectangles) with 7.5 mM MβCD for 30 min. MβCD was washed out, and various amounts of cholesterol in MβCD were added. After 30 min, the cholesterol was removed, and the cells were infected with HSV-1 (KOS/tk12) and assayed for β-galactosidase activity at 6 h postinfection. The results presented are representative of two independent experiments done in triplicate.

    Techniques Used: Infection, Activity Assay

    3) Product Images from "Targeting an Oncolytic Influenza A Virus to Tumor Tissue by Elastase"

    Article Title: Targeting an Oncolytic Influenza A Virus to Tumor Tissue by Elastase

    Journal: Molecular Therapy Oncolytics

    doi: 10.1016/j.omto.2017.09.002

    Growth of NS 116 -GFP/AT and NS 116 -GFP/AE Viruses in Different Cell Lines Vero, B16f1, PANC-1, A375, or CaCo2 1 × 10 5 cells were infected with either NS 116 -GFP/AT or NS 116 -GFP/AE viruses at an MOI of 0.01 and cultivated in the presence trypsin or neutrophil elastase, respectively. Supernatant was collected 12, 24, 48, and 72 hr post-infection. Data represent means + SEM (n = 3).
    Figure Legend Snippet: Growth of NS 116 -GFP/AT and NS 116 -GFP/AE Viruses in Different Cell Lines Vero, B16f1, PANC-1, A375, or CaCo2 1 × 10 5 cells were infected with either NS 116 -GFP/AT or NS 116 -GFP/AE viruses at an MOI of 0.01 and cultivated in the presence trypsin or neutrophil elastase, respectively. Supernatant was collected 12, 24, 48, and 72 hr post-infection. Data represent means + SEM (n = 3).

    Techniques Used: Infection

    Growth Kinetic of Modified NS 116 -GFP/AE Virus A 24-hr-old monolayer of Vero cells was infected with NS 116 -GFP/AE virus in the presence of either pancreatic elastase (p-elastase) or neutrophil elastase (n-elastase). A viral titer was determined by TCID 50 in Vero cells at the indicated time points + SEM (n = 6).
    Figure Legend Snippet: Growth Kinetic of Modified NS 116 -GFP/AE Virus A 24-hr-old monolayer of Vero cells was infected with NS 116 -GFP/AE virus in the presence of either pancreatic elastase (p-elastase) or neutrophil elastase (n-elastase). A viral titer was determined by TCID 50 in Vero cells at the indicated time points + SEM (n = 6).

    Techniques Used: Modification, Infection

    Growth Properties of NS 116 -GFP/AE and NS 116 -GFP/AT Virus in Vero Cells in Co-culture System with Human Neutrophils or Supernatant Supplemented with Neutrophil Elastase or Trypsin (A) Vero cells were infected at an MOI of 0.01 under indicated condition and viral titer was determined by TCID 50 24 hr post-infection. Data represent means + SEM (n = 3). (B) Virus induced GFP expression, which indicates viral growth. The pictures of infected Vero cells were taken 48 hr post-infection using an AxioCam ICc3 Rev.2-3 camera, with a magnification of 200×. Experiments were carried out three times with neutrophils of another proband, and the corresponding results are provided. Scale bar, 100 μm.
    Figure Legend Snippet: Growth Properties of NS 116 -GFP/AE and NS 116 -GFP/AT Virus in Vero Cells in Co-culture System with Human Neutrophils or Supernatant Supplemented with Neutrophil Elastase or Trypsin (A) Vero cells were infected at an MOI of 0.01 under indicated condition and viral titer was determined by TCID 50 24 hr post-infection. Data represent means + SEM (n = 3). (B) Virus induced GFP expression, which indicates viral growth. The pictures of infected Vero cells were taken 48 hr post-infection using an AxioCam ICc3 Rev.2-3 camera, with a magnification of 200×. Experiments were carried out three times with neutrophils of another proband, and the corresponding results are provided. Scale bar, 100 μm.

    Techniques Used: Co-Culture Assay, Infection, Expressing

    4) Product Images from "Mucin-Like Domain of Ebola Virus Glycoprotein Enhances Selective Oncolytic Actions against Brain Tumors"

    Article Title: Mucin-Like Domain of Ebola Virus Glycoprotein Enhances Selective Oncolytic Actions against Brain Tumors

    Journal: Journal of Virology

    doi: 10.1128/JVI.01967-19

    Glioblastoma cultures yield larger plaques than normal human glia. (A) Representative images of viral plaques at 2 dpi after fixation and fluorescent immunolabeling in cultures of human glia, human glioblastoma (ATCC87, A172, U1118, U251), and mouse glioma (CT-2A) cells. Three chimeric VSV-EBOVs (VSV-EBOV, VSV-EBOVΔMLD, VSV-EBOVΔMLD-GFP) and the attenuated nonchimeric VSV (VSVwt-GFP) were tested. Bar, 1 mm. (B) Graphs showing viral plaque size development over time. Parallel cultures of the cells shown in panel A were infected and overlaid in a plaque assay and then allowed to develop for 1, 2, or 3 days prior to fixation and immunolabeling. Plaques were visualized using fluorescence microscopy, and an area of 60 plaques was measured for each condition (mean ± SE). (C) One-step growth curves comparing VSV-EBOV (black) and VSV-EBOVΔMLD (red) performed on cultures of mouse primary brain, human glioma ATCC87, and Vero cells. Each curve represents the mean from duplicate experiments, with the titers at the indicated time points being determined by plaque assay on Vero cells. (D) Summary bar graph of one-step viral titers from panel C measured at 10 h. (E) Bar graph comparing the relative levels of intracellular VSV N RNA from VSV-EBOV- and VSV-EBOVΔMLD-infected Vero cell cultures (MOI = 10). RNA was harvested from cells at 3, 6, and 12 h postinfection, and VSV RNA was measured using RT-qPCR. (F) SDS-PAGE Western blots comparing the relative amounts of intracellular VSV N and M proteins using conditions similar to those used for the RNA analysis for which the results are shown in panel E. Results from triplicate infections are shown. **, P
    Figure Legend Snippet: Glioblastoma cultures yield larger plaques than normal human glia. (A) Representative images of viral plaques at 2 dpi after fixation and fluorescent immunolabeling in cultures of human glia, human glioblastoma (ATCC87, A172, U1118, U251), and mouse glioma (CT-2A) cells. Three chimeric VSV-EBOVs (VSV-EBOV, VSV-EBOVΔMLD, VSV-EBOVΔMLD-GFP) and the attenuated nonchimeric VSV (VSVwt-GFP) were tested. Bar, 1 mm. (B) Graphs showing viral plaque size development over time. Parallel cultures of the cells shown in panel A were infected and overlaid in a plaque assay and then allowed to develop for 1, 2, or 3 days prior to fixation and immunolabeling. Plaques were visualized using fluorescence microscopy, and an area of 60 plaques was measured for each condition (mean ± SE). (C) One-step growth curves comparing VSV-EBOV (black) and VSV-EBOVΔMLD (red) performed on cultures of mouse primary brain, human glioma ATCC87, and Vero cells. Each curve represents the mean from duplicate experiments, with the titers at the indicated time points being determined by plaque assay on Vero cells. (D) Summary bar graph of one-step viral titers from panel C measured at 10 h. (E) Bar graph comparing the relative levels of intracellular VSV N RNA from VSV-EBOV- and VSV-EBOVΔMLD-infected Vero cell cultures (MOI = 10). RNA was harvested from cells at 3, 6, and 12 h postinfection, and VSV RNA was measured using RT-qPCR. (F) SDS-PAGE Western blots comparing the relative amounts of intracellular VSV N and M proteins using conditions similar to those used for the RNA analysis for which the results are shown in panel E. Results from triplicate infections are shown. **, P

    Techniques Used: Immunolabeling, Infection, Plaque Assay, Fluorescence, Microscopy, Quantitative RT-PCR, SDS Page, Western Blot

    5) Product Images from "The Cell Cycle and Virus Infection"

    Article Title: The Cell Cycle and Virus Infection

    Journal: Cell Cycle Control

    doi: 10.1385/1-59259-857-9:197

    Localization of caspase 8 in transfected or etoposide-treated Vero cells or cells expressing the avian coronavirus nucleoprotein. Vero cells were transfected with pTriExIB VNHis (a construct that expresses the nucleoprotein under the control of a PolII promoter), treated with 100 mM etoposide, or left untreated and analyzed by indirect immunofluorescence for the localization of caspse 8 using a polyclonal anticaspase 8 (P 20) antibody (Santa Cruz Biotechnology). Transfected cells were analyzed 24 h post transfection, whereas etoposide-treated cells were analyzed 48 h post treatment. In nontreated cells (left row) caspase 8 can be detected in both the nucleus and the cytoplasm, in cells transfected with IB VN His (middle row), caspase 8 is localised in the cyto-and nucleoplasm with a prominent signal in the perinuclear region and the nucleolus. In Etoposide treated cells (right row) caspse 8 is almost exclusively localised in the nucleus and the perinuclear region. Primary caspase 8 antibody was detected with FITCconjugated goat antirabbit antibody (Sigma), and IBV NHis was detected with mouse anti-His (C-term; Sigma) antibody as primary antibodies and Texas red-conjugated goat anti mouse (Harlan Sera Lab) as secondary antibody.
    Figure Legend Snippet: Localization of caspase 8 in transfected or etoposide-treated Vero cells or cells expressing the avian coronavirus nucleoprotein. Vero cells were transfected with pTriExIB VNHis (a construct that expresses the nucleoprotein under the control of a PolII promoter), treated with 100 mM etoposide, or left untreated and analyzed by indirect immunofluorescence for the localization of caspse 8 using a polyclonal anticaspase 8 (P 20) antibody (Santa Cruz Biotechnology). Transfected cells were analyzed 24 h post transfection, whereas etoposide-treated cells were analyzed 48 h post treatment. In nontreated cells (left row) caspase 8 can be detected in both the nucleus and the cytoplasm, in cells transfected with IB VN His (middle row), caspase 8 is localised in the cyto-and nucleoplasm with a prominent signal in the perinuclear region and the nucleolus. In Etoposide treated cells (right row) caspse 8 is almost exclusively localised in the nucleus and the perinuclear region. Primary caspase 8 antibody was detected with FITCconjugated goat antirabbit antibody (Sigma), and IBV NHis was detected with mouse anti-His (C-term; Sigma) antibody as primary antibodies and Texas red-conjugated goat anti mouse (Harlan Sera Lab) as secondary antibody.

    Techniques Used: Transfection, Expressing, Construct, Immunofluorescence

    (A) Flow cytometry analysis of mock and avian coronavirus-infected cells at 0 and 16 h post infection. (B) Detection of avian coronavirus protein by indirect immunofluorescence in infected cells. Vero cells were infected with coronavirus for 0 h or 16 h, fixed, and analyzed by indirect immunoflourescence using appropriate antibodies. (Original magnification×620.) The data indicate that infected cells accumulate in S-phase compared with mock infected cells.
    Figure Legend Snippet: (A) Flow cytometry analysis of mock and avian coronavirus-infected cells at 0 and 16 h post infection. (B) Detection of avian coronavirus protein by indirect immunofluorescence in infected cells. Vero cells were infected with coronavirus for 0 h or 16 h, fixed, and analyzed by indirect immunoflourescence using appropriate antibodies. (Original magnification×620.) The data indicate that infected cells accumulate in S-phase compared with mock infected cells.

    Techniques Used: Flow Cytometry, Infection, Immunofluorescence

    6) Product Images from "Ribavirin efficiently suppresses porcine nidovirus replication"

    Article Title: Ribavirin efficiently suppresses porcine nidovirus replication

    Journal: Virus Research

    doi: 10.1016/j.virusres.2012.10.018

    Inhibition of viral protein translation by ribavirin. Ribavirin-treated PAM-pCD163 and Vero cells were mock-infected or infected with PRRSV (A) or PEDV (B) for 1 h and were further cultivated in the presence or absence of ribavirin. At 48 hpi, cellular lysates were collected, resolved by SDS-PAGE, transferred to a nitrocellulose membrane, and immunoblotted by using the antibody that recognizes the PRRSV N protein or the PEDV S protein. The blot was also reacted with mouse MAb against β-actin to verify equal protein loading. Each viral protein expression was quantitatively analyzed by densitometry in terms of the relative density value to the β-actin gene and ribavirin-treated sample results were compared to vehicle-control results. Values are representative of the mean from three independent experiments and error bars denote standard deviations. * P = 0.001–0.05; † P
    Figure Legend Snippet: Inhibition of viral protein translation by ribavirin. Ribavirin-treated PAM-pCD163 and Vero cells were mock-infected or infected with PRRSV (A) or PEDV (B) for 1 h and were further cultivated in the presence or absence of ribavirin. At 48 hpi, cellular lysates were collected, resolved by SDS-PAGE, transferred to a nitrocellulose membrane, and immunoblotted by using the antibody that recognizes the PRRSV N protein or the PEDV S protein. The blot was also reacted with mouse MAb against β-actin to verify equal protein loading. Each viral protein expression was quantitatively analyzed by densitometry in terms of the relative density value to the β-actin gene and ribavirin-treated sample results were compared to vehicle-control results. Values are representative of the mean from three independent experiments and error bars denote standard deviations. * P = 0.001–0.05; † P

    Techniques Used: Inhibition, Infection, SDS Page, Expressing

    Suppression of nidovirus propagation by ribavirin at early times post-infection. PAM-pCD163 and Vero cells were pretreated with ribavirin and were infected with PRRSV (A) or PEDV (B), respectively. At the indicated times at post-infection, ribavirin was added to achieve a final concentration. At 48 hpi, virus-infected cells were fixed and virus infectivity was determined by quantifying the number of cells expressing N proteins through IFA. Data are representative of the mean of three independent experiments and error bars represent standard deviations. * P = 0.001–0.05; † P
    Figure Legend Snippet: Suppression of nidovirus propagation by ribavirin at early times post-infection. PAM-pCD163 and Vero cells were pretreated with ribavirin and were infected with PRRSV (A) or PEDV (B), respectively. At the indicated times at post-infection, ribavirin was added to achieve a final concentration. At 48 hpi, virus-infected cells were fixed and virus infectivity was determined by quantifying the number of cells expressing N proteins through IFA. Data are representative of the mean of three independent experiments and error bars represent standard deviations. * P = 0.001–0.05; † P

    Techniques Used: Infection, Concentration Assay, Expressing, Immunofluorescence

    Reduction of viral progeny production by ribavirin. (A) PAM-pCD163 and Vero cells were pretreated with ribavirin for 1 h and were mock or virus infected (MOI of 0.1). Ribavirin was present in the medium throughout the infection. At 48 hpi, the virus supernatants were collected and the titers of PRRSV (left) and PEDV (right) were determined. (B) Growth kinetics of PRRSV (left) and PEDV (right) upon treatment with ribavirin was assessed exactly as for panel A. At the indicated time points post-infection, culture supernatants were harvested and virus titers were measured. (C) Virus internalization assay. PAM-pCD163 and Vero cells were infected at an MOI of 0.1 at 4 °C for 1 h. After washing with cold PBS, infected cells were maintained in the presence or absence of ribavirin either at 4 °C or 37 °C for an additional hour. Bound but uninternalized virus particles were removed by treatment with protease K. The infected cells were then serially diluted and plated onto fresh target cells. At 2 days post-incubation, internalized viruses were titrated by IFA and plaque assay for PRRSV (left) and PEDV (right), respectively. Results are expressed as the mean values from triplicate wells and error bars represent standard deviations. * P = 0.001–0.05; † P
    Figure Legend Snippet: Reduction of viral progeny production by ribavirin. (A) PAM-pCD163 and Vero cells were pretreated with ribavirin for 1 h and were mock or virus infected (MOI of 0.1). Ribavirin was present in the medium throughout the infection. At 48 hpi, the virus supernatants were collected and the titers of PRRSV (left) and PEDV (right) were determined. (B) Growth kinetics of PRRSV (left) and PEDV (right) upon treatment with ribavirin was assessed exactly as for panel A. At the indicated time points post-infection, culture supernatants were harvested and virus titers were measured. (C) Virus internalization assay. PAM-pCD163 and Vero cells were infected at an MOI of 0.1 at 4 °C for 1 h. After washing with cold PBS, infected cells were maintained in the presence or absence of ribavirin either at 4 °C or 37 °C for an additional hour. Bound but uninternalized virus particles were removed by treatment with protease K. The infected cells were then serially diluted and plated onto fresh target cells. At 2 days post-incubation, internalized viruses were titrated by IFA and plaque assay for PRRSV (left) and PEDV (right), respectively. Results are expressed as the mean values from triplicate wells and error bars represent standard deviations. * P = 0.001–0.05; † P

    Techniques Used: Infection, Incubation, Immunofluorescence, Plaque Assay

    Effect of ribavirin on the replication of porcine nidoviruses. PAM-pCD163 and Vero cells were preincubated with ribavirin at indicated concentrations for 1 h prior to infection and were mock-infected or infected with PRRSV (A) or PEDV (B) at an MOI of 0.1. Virus-infected cells were further maintained for 48 h in the presence of vehicle or ribavirin. Virus-specific CPEs were observed daily and were photographed at 48 hpi using an inverted microscope at a magnification of 100× (first panels). For immunostaining, infected cells were fixed at 48 hpi and incubated with MAb against the N protein of PRRSV or PEDV followed by Alexa green-conjugated goat anti-mouse secondary antibody (second panels). The cells were then counterstained with DAPI (third panels) and examined using a fluorescent microscope at 200× magnification. Viral productions in the presence of ribavirin were measured by quantifying the number of cells expressing N proteins through IFA. Five fields at 200× magnification were counted per each condition and the total number of cells per field as determined by DAPI staining was similar in all fields. Values are representative of the mean of three independent experiments and error bars represent standard deviations. * P = 0.001–0.05; † P
    Figure Legend Snippet: Effect of ribavirin on the replication of porcine nidoviruses. PAM-pCD163 and Vero cells were preincubated with ribavirin at indicated concentrations for 1 h prior to infection and were mock-infected or infected with PRRSV (A) or PEDV (B) at an MOI of 0.1. Virus-infected cells were further maintained for 48 h in the presence of vehicle or ribavirin. Virus-specific CPEs were observed daily and were photographed at 48 hpi using an inverted microscope at a magnification of 100× (first panels). For immunostaining, infected cells were fixed at 48 hpi and incubated with MAb against the N protein of PRRSV or PEDV followed by Alexa green-conjugated goat anti-mouse secondary antibody (second panels). The cells were then counterstained with DAPI (third panels) and examined using a fluorescent microscope at 200× magnification. Viral productions in the presence of ribavirin were measured by quantifying the number of cells expressing N proteins through IFA. Five fields at 200× magnification were counted per each condition and the total number of cells per field as determined by DAPI staining was similar in all fields. Values are representative of the mean of three independent experiments and error bars represent standard deviations. * P = 0.001–0.05; † P

    Techniques Used: Infection, Inverted Microscopy, Immunostaining, Incubation, Microscopy, Expressing, Immunofluorescence, Staining

    Inhibition of viral RNA transcription by ribavirin. PAM-pCD163 and Vero cells pretreated with ribavirin were mock-infected or infected with PRRSV (A) or PEDV (B) for 1 h and were incubated in the presence of ribavirin. Total cellular RNA was extracted at 48 hpi, and strand-specific viral genomic RNAs (black bars) and sg mRNAs (white bars) of PRRSV and PEDV were amplified by quantitative real-time RT-PCR. Viral positive-sense genomic RNA and sg mRNA were normalized to mRNA for porcine β-actin or monkey GAPDH and relative quantities (RQ) of mRNA accumulation were evaluated. Ribavirin-treated sample results were compared with untreated results. Values are representative of the mean from three independent experiments and error bars denote standard deviations. * P = 0.001–0.05; † P
    Figure Legend Snippet: Inhibition of viral RNA transcription by ribavirin. PAM-pCD163 and Vero cells pretreated with ribavirin were mock-infected or infected with PRRSV (A) or PEDV (B) for 1 h and were incubated in the presence of ribavirin. Total cellular RNA was extracted at 48 hpi, and strand-specific viral genomic RNAs (black bars) and sg mRNAs (white bars) of PRRSV and PEDV were amplified by quantitative real-time RT-PCR. Viral positive-sense genomic RNA and sg mRNA were normalized to mRNA for porcine β-actin or monkey GAPDH and relative quantities (RQ) of mRNA accumulation were evaluated. Ribavirin-treated sample results were compared with untreated results. Values are representative of the mean from three independent experiments and error bars denote standard deviations. * P = 0.001–0.05; † P

    Techniques Used: Inhibition, Infection, Incubation, Amplification, Quantitative RT-PCR

    Effects of supplementation of guanosine in ribavirin treatment and MPA on porcine nidovirus infection. (A) Effect of ribavirin alone or with guanosine (100 μM) on the replication of PRRSV (left) and PEDV (right). PAM-pCD163 and Vero cells were preincubated with various concentrations of ribavirin with (+) or without 100 (−) μM guanosine. Virus-infected cells were fixed at 48 hpi and subjected to immunofluorescence assay. Viral productions were measured by quantifying the number of cells expressing N proteins through IFA as described above. Viral productions in cells treated by guanosine and ribavirin that were significantly different from those in cells treated with ribavirin alone are indicated. (B) Effect of MPA on the replication of PRRSV (left) and PEDV (right). PAM-pCD163 and Vero cells were preincubated with various concentrations of MPA ranging from 0 (mock) to 10 μM and infected with PRRSV or PEDV. At 48 hpi, virus-infected cells were subjected to immunofluorescence assay, and viral productions were measured by quantifying the number of cells expressing N proteins through IFA as described above. Data are representative of the mean values from three independent experiments and error bars represent standard deviations. * P = 0.001–0.05; † P
    Figure Legend Snippet: Effects of supplementation of guanosine in ribavirin treatment and MPA on porcine nidovirus infection. (A) Effect of ribavirin alone or with guanosine (100 μM) on the replication of PRRSV (left) and PEDV (right). PAM-pCD163 and Vero cells were preincubated with various concentrations of ribavirin with (+) or without 100 (−) μM guanosine. Virus-infected cells were fixed at 48 hpi and subjected to immunofluorescence assay. Viral productions were measured by quantifying the number of cells expressing N proteins through IFA as described above. Viral productions in cells treated by guanosine and ribavirin that were significantly different from those in cells treated with ribavirin alone are indicated. (B) Effect of MPA on the replication of PRRSV (left) and PEDV (right). PAM-pCD163 and Vero cells were preincubated with various concentrations of MPA ranging from 0 (mock) to 10 μM and infected with PRRSV or PEDV. At 48 hpi, virus-infected cells were subjected to immunofluorescence assay, and viral productions were measured by quantifying the number of cells expressing N proteins through IFA as described above. Data are representative of the mean values from three independent experiments and error bars represent standard deviations. * P = 0.001–0.05; † P

    Techniques Used: Infection, Immunofluorescence, Expressing

    7) Product Images from "Antiviral and Antioxidant Properties of Echinochrome A"

    Article Title: Antiviral and Antioxidant Properties of Echinochrome A

    Journal: Marine Drugs

    doi: 10.3390/md16120509

    Influence of Ech and studied formulations on the lipopolysaccharide (LPS)-induced reactive oxygen species (ROS) formations in Vero cells. The formulations were tested at a concentration of five μg/mL. * p
    Figure Legend Snippet: Influence of Ech and studied formulations on the lipopolysaccharide (LPS)-induced reactive oxygen species (ROS) formations in Vero cells. The formulations were tested at a concentration of five μg/mL. * p

    Techniques Used: Concentration Assay

    8) Product Images from "Real-time monitoring of adherent Vero cell density and apoptosis in bioreactor processes"

    Article Title: Real-time monitoring of adherent Vero cell density and apoptosis in bioreactor processes

    Journal: Cytotechnology

    doi: 10.1007/s10616-011-9421-2

    Microscopic observation of Vero cells attached on microcarriers. Evolution of Vero cell morphology on microporous microcarriers at 4 h ( A ), 56 h ( B ), 70 h ( C ) and 94 h ( D ) after cell seeding, during culture performed with
    Figure Legend Snippet: Microscopic observation of Vero cells attached on microcarriers. Evolution of Vero cell morphology on microporous microcarriers at 4 h ( A ), 56 h ( B ), 70 h ( C ) and 94 h ( D ) after cell seeding, during culture performed with

    Techniques Used:

    Evolution of the permittivity and the specific permittivity, ε X , with the adhered Vero cell concentration. A Correlation between permittivity and concentration of Vero cells attached on microcarriers, during Batch 1 (B1: black circle ), Batch
    Figure Legend Snippet: Evolution of the permittivity and the specific permittivity, ε X , with the adhered Vero cell concentration. A Correlation between permittivity and concentration of Vero cells attached on microcarriers, during Batch 1 (B1: black circle ), Batch

    Techniques Used: Concentration Assay

    9) Product Images from "Development of improved vaccine cell lines against rotavirus"

    Article Title: Development of improved vaccine cell lines against rotavirus

    Journal: Scientific Data

    doi: 10.1038/sdata.2017.21

    Experimental workflow. This study included a tiered siRNA screening approach coupled with CRISPR-generated knockout Vero cell lines to evaluate host genes and their implication during RV infection. The primary siRNA screen was performed with pooled OTP-siRNA in MA104 cell line using the simian rotavirus strain RV3. Top hits were selected by a z-score analysis and subjected to deconvoluted validation siRNA screens in Vero cells with the rotavirus strains RV3-BB, Rotarix, CDC-9. Resulting hits were evaluated in CRISPR generated Vero cell lines with CDC-9 and Rotarix (GSK P5) strains.
    Figure Legend Snippet: Experimental workflow. This study included a tiered siRNA screening approach coupled with CRISPR-generated knockout Vero cell lines to evaluate host genes and their implication during RV infection. The primary siRNA screen was performed with pooled OTP-siRNA in MA104 cell line using the simian rotavirus strain RV3. Top hits were selected by a z-score analysis and subjected to deconvoluted validation siRNA screens in Vero cells with the rotavirus strains RV3-BB, Rotarix, CDC-9. Resulting hits were evaluated in CRISPR generated Vero cell lines with CDC-9 and Rotarix (GSK P5) strains.

    Techniques Used: CRISPR, Generated, Knock-Out, Infection

    10) Product Images from "Machupo Virus Expressing GPC of the Candid#1 Vaccine Strain of Junin Virus Is Highly Attenuated and Immunogenic"

    Article Title: Machupo Virus Expressing GPC of the Candid#1 Vaccine Strain of Junin Virus Is Highly Attenuated and Immunogenic

    Journal: Journal of Virology

    doi: 10.1128/JVI.02615-15

    Schematic representation of the genome of rMACV/Cd#1-GPC and the virus growth curves. (A) For rMACV/Cd#1-GPC, the entire MACV GPC gene was replaced with the Cd#1 GPC gene. The plaque morphology is shown. (B) The growth of rMACV/Cd#1-GPC was characterized in Vero cells (MOI = 0.01). The titer of rMACV was significantly higher than that of rCd#1 at 48 hpi ( n = 5; **, P
    Figure Legend Snippet: Schematic representation of the genome of rMACV/Cd#1-GPC and the virus growth curves. (A) For rMACV/Cd#1-GPC, the entire MACV GPC gene was replaced with the Cd#1 GPC gene. The plaque morphology is shown. (B) The growth of rMACV/Cd#1-GPC was characterized in Vero cells (MOI = 0.01). The titer of rMACV was significantly higher than that of rCd#1 at 48 hpi ( n = 5; **, P

    Techniques Used: Gel Permeation Chromatography

    11) Product Images from "Mutant Fusion Proteins with Enhanced Fusion Activity Promote Measles Virus Spread in Human Neuronal Cells and Brains of Suckling Hamsters"

    Article Title: Mutant Fusion Proteins with Enhanced Fusion Activity Promote Measles Virus Spread in Human Neuronal Cells and Brains of Suckling Hamsters

    Journal: Journal of Virology

    doi: 10.1128/JVI.02632-12

    Infection with the recombinant MV with the F protein lacking the cytoplasmic tail (IC323-EGFP-FΔ30). SLAM- and nectin 4-negative Vero (A) and IMR-32 (B) cells were infected with IC323-EGFP or IC323-EGFP-FΔ30 at an MOI of 0.1. At 72 h after
    Figure Legend Snippet: Infection with the recombinant MV with the F protein lacking the cytoplasmic tail (IC323-EGFP-FΔ30). SLAM- and nectin 4-negative Vero (A) and IMR-32 (B) cells were infected with IC323-EGFP or IC323-EGFP-FΔ30 at an MOI of 0.1. At 72 h after

    Techniques Used: Infection, Recombinant

    Amino acid substitutions in the F protein enhance cell-cell fusion in Vero/hSLAM and Vero cells. (A) Vero/hSLAM cells were transfected with expression plasmids encoding wild-type (Wt) or mutant F proteins with the indicated substitutions, together with
    Figure Legend Snippet: Amino acid substitutions in the F protein enhance cell-cell fusion in Vero/hSLAM and Vero cells. (A) Vero/hSLAM cells were transfected with expression plasmids encoding wild-type (Wt) or mutant F proteins with the indicated substitutions, together with

    Techniques Used: Transfection, Expressing, Mutagenesis

    Syncytium formation in SLAM- and nectin 4-negative cells infected with recombinant MVs. Vero (A), IMR-32 (B), and SYM-1 (C) cells were infected with the parental recombinant MV expressing EGFP (IC323-EGFP) or its mutants expressing the F protein with
    Figure Legend Snippet: Syncytium formation in SLAM- and nectin 4-negative cells infected with recombinant MVs. Vero (A), IMR-32 (B), and SYM-1 (C) cells were infected with the parental recombinant MV expressing EGFP (IC323-EGFP) or its mutants expressing the F protein with

    Techniques Used: Infection, Recombinant, Expressing

    12) Product Images from "Binding of DC-SIGN to the Hemagglutinin of Influenza A Viruses Supports Virus Replication in DC-SIGN Expressing Cells"

    Article Title: Binding of DC-SIGN to the Hemagglutinin of Influenza A Viruses Supports Virus Replication in DC-SIGN Expressing Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0056164

    The number of glycosylation sites present on HA determines the virus infection rates in DC-SIGN expressing cells. MDCK (A) and Vero (B) cells, transfected with the DC-SIGN gene (black bars) or not (white bars) were treated with neuraminidase from vibrio cholerae and GolgiStop for 30 minutes to remove sialic acids from the cell surface. These cells were subsequently inoculated with A/Netherlands/26/07, A/Netherlands/26/07-Δ125, A/Netherlands/602/09, A/Netherlands/602/09-Δ276 or A/Netherlands/602/09-VN54 N125 N160. The percentage of infected cells relative to the positive control (untreated cells still possessing sialic acid) was assessed after detecting infected cells using a FITC-labeled antibody to the viral nucleoprotein and flow-cytometry.
    Figure Legend Snippet: The number of glycosylation sites present on HA determines the virus infection rates in DC-SIGN expressing cells. MDCK (A) and Vero (B) cells, transfected with the DC-SIGN gene (black bars) or not (white bars) were treated with neuraminidase from vibrio cholerae and GolgiStop for 30 minutes to remove sialic acids from the cell surface. These cells were subsequently inoculated with A/Netherlands/26/07, A/Netherlands/26/07-Δ125, A/Netherlands/602/09, A/Netherlands/602/09-Δ276 or A/Netherlands/602/09-VN54 N125 N160. The percentage of infected cells relative to the positive control (untreated cells still possessing sialic acid) was assessed after detecting infected cells using a FITC-labeled antibody to the viral nucleoprotein and flow-cytometry.

    Techniques Used: Infection, Expressing, Transfection, Positive Control, Labeling, Flow Cytometry, Cytometry

    Expression of DC-SIGN supports replication of influenza A viruses in the absence of sialic acids. MDCK (A and C) and Vero cells (B and D) transfected with the DC-SIGN gene (black bars) or not (white bars), were treated with neuraminidase from vibrio cholerae and GolgiStop for 30 minutes to remove sialic acids from the cell surface. These cells were subsequently inoculated with five different A/H1N1 viruses (A and B) and four A/H3N2 viruses (C and D). The percentage infected cells relative to the untreated control cells, still possessing sialic acid, was assessed after detecting infected cells using a FITC-labelled antibody to the viral nucleoprotein and flow-cytometry. To confirm that the entry was mediated via DC-SIGN, Vero and Vero DC-SIGN were treated with neuraminidase from vibrio cholerae for 30 minutes to remove sialic acids from the cell surface and incubated with or without antibodies to DC-SIGN or an isotype control antibody as indicated (E and F). These cells were subsequently inoculated with influenza viruses. NL/312/03 and USSR/90/77. The percentage of infected cells compared to the positive control (untreated cells, still possessing sialic acid) was assessed as described above.
    Figure Legend Snippet: Expression of DC-SIGN supports replication of influenza A viruses in the absence of sialic acids. MDCK (A and C) and Vero cells (B and D) transfected with the DC-SIGN gene (black bars) or not (white bars), were treated with neuraminidase from vibrio cholerae and GolgiStop for 30 minutes to remove sialic acids from the cell surface. These cells were subsequently inoculated with five different A/H1N1 viruses (A and B) and four A/H3N2 viruses (C and D). The percentage infected cells relative to the untreated control cells, still possessing sialic acid, was assessed after detecting infected cells using a FITC-labelled antibody to the viral nucleoprotein and flow-cytometry. To confirm that the entry was mediated via DC-SIGN, Vero and Vero DC-SIGN were treated with neuraminidase from vibrio cholerae for 30 minutes to remove sialic acids from the cell surface and incubated with or without antibodies to DC-SIGN or an isotype control antibody as indicated (E and F). These cells were subsequently inoculated with influenza viruses. NL/312/03 and USSR/90/77. The percentage of infected cells compared to the positive control (untreated cells, still possessing sialic acid) was assessed as described above.

    Techniques Used: Expressing, Transfection, Infection, Flow Cytometry, Cytometry, Incubation, Positive Control

    DC-SIGN expression in stably transfected MDCK DC-SIGN and Vero DC-SIGN cells. MDCK cells (A) and Vero cells (B) without (dotted line) and transfected with the gene encoding DC-SIGN (solid line) were analyzed for DC-SIGN expression after staining with a PE-labeled antibody to DC-SIGN and flow cytometry.
    Figure Legend Snippet: DC-SIGN expression in stably transfected MDCK DC-SIGN and Vero DC-SIGN cells. MDCK cells (A) and Vero cells (B) without (dotted line) and transfected with the gene encoding DC-SIGN (solid line) were analyzed for DC-SIGN expression after staining with a PE-labeled antibody to DC-SIGN and flow cytometry.

    Techniques Used: Expressing, Stable Transfection, Transfection, Staining, Labeling, Flow Cytometry, Cytometry

    13) Product Images from "Expression of an Epitope-Tagged Virulence Protein in Rickettsia parkeri Using Transposon Insertion"

    Article Title: Expression of an Epitope-Tagged Virulence Protein in Rickettsia parkeri Using Transposon Insertion

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0037310

    Detection of FLAG-RickA in bacteria. R. parkeri strains that were not transformed ( Rp ) or transformed with pMW1650-FLAG-RickA ( Rp FLAG-RickA) were used to infect Vero cells and then (A) labeled by immunofluorescence with anti-RickA antibody and stained for DNA with DAPI, or (B) labeled by immunofluorescence with anti-FLAG antibody and stained for DNA with DAPI. In the merged images, RickA or FLAG are labeled in green, and DNA in red. Scale bar 10 µm. Higher magnification images of individual bacteria (highlighted in boxes in the lower magnification images) are shown on the right.
    Figure Legend Snippet: Detection of FLAG-RickA in bacteria. R. parkeri strains that were not transformed ( Rp ) or transformed with pMW1650-FLAG-RickA ( Rp FLAG-RickA) were used to infect Vero cells and then (A) labeled by immunofluorescence with anti-RickA antibody and stained for DNA with DAPI, or (B) labeled by immunofluorescence with anti-FLAG antibody and stained for DNA with DAPI. In the merged images, RickA or FLAG are labeled in green, and DNA in red. Scale bar 10 µm. Higher magnification images of individual bacteria (highlighted in boxes in the lower magnification images) are shown on the right.

    Techniques Used: Transformation Assay, Labeling, Immunofluorescence, Staining

    14) Product Images from "A Carbohydrate Moiety of Secreted Stage-Specific Glycoprotein 4 Participates in Host Cell Invasion by Trypanosoma cruzi Extracellular Amastigotes"

    Article Title: A Carbohydrate Moiety of Secreted Stage-Specific Glycoprotein 4 Participates in Host Cell Invasion by Trypanosoma cruzi Extracellular Amastigotes

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2018.00693

    Extracellular amastigotes release Ssp-4 associated with vesicles interacting with host cell. (A) Scanning electron microscopy of extracellular amastigotes (EAs) adhered to coverslips coated with poly-L-lysine or Vero cells. Scale bars: 2 and 5 μm. (B) Transmission electron microscopy of HeLa cells incubated for 30 min with EAs from the G strain (colored in red). Scale bar: 1 μm. Black arrows indicate EA secreted vesicles. (C) Immunofluorescence of HeLa cells incubated for 30 min with EAs from G (MOI 10:1) and mAb1D9 (green), DAPI (blue), and phalloidin-TRITC (red). Upper panels: images of a focal plane showing vesicles secreted by EAs of the G strain (green) associated with actin (red); bottom panels: three-dimensional reconstruction from a Z -series acquired by a confocal microscope. Scale bar: 2 μm. Arrows indicate vesicular structures secreted by EAs. (D) Extracellular amastigotes (EAs) from the G and Y strains were incubated for 6 h in RPMI medium without fetal bovine serum. Supernatant was collected and fractionated in three different populations: V2 (pelleted after 2 h of ultracentrifugation), V16 (pelleted after 16 h ultracentrifugation), and VF (supernatant from pellet V16). Western blotting with mAb1D9 and silver-stained SDS–PAGE of the different fractions. Whole cell (WC) lysate of EAs of the G strain was used as control.
    Figure Legend Snippet: Extracellular amastigotes release Ssp-4 associated with vesicles interacting with host cell. (A) Scanning electron microscopy of extracellular amastigotes (EAs) adhered to coverslips coated with poly-L-lysine or Vero cells. Scale bars: 2 and 5 μm. (B) Transmission electron microscopy of HeLa cells incubated for 30 min with EAs from the G strain (colored in red). Scale bar: 1 μm. Black arrows indicate EA secreted vesicles. (C) Immunofluorescence of HeLa cells incubated for 30 min with EAs from G (MOI 10:1) and mAb1D9 (green), DAPI (blue), and phalloidin-TRITC (red). Upper panels: images of a focal plane showing vesicles secreted by EAs of the G strain (green) associated with actin (red); bottom panels: three-dimensional reconstruction from a Z -series acquired by a confocal microscope. Scale bar: 2 μm. Arrows indicate vesicular structures secreted by EAs. (D) Extracellular amastigotes (EAs) from the G and Y strains were incubated for 6 h in RPMI medium without fetal bovine serum. Supernatant was collected and fractionated in three different populations: V2 (pelleted after 2 h of ultracentrifugation), V16 (pelleted after 16 h ultracentrifugation), and VF (supernatant from pellet V16). Western blotting with mAb1D9 and silver-stained SDS–PAGE of the different fractions. Whole cell (WC) lysate of EAs of the G strain was used as control.

    Techniques Used: Electron Microscopy, Transmission Assay, Incubation, Immunofluorescence, Microscopy, Western Blot, Staining, SDS Page

    15) Product Images from "A Carbohydrate Moiety of Secreted Stage-Specific Glycoprotein 4 Participates in Host Cell Invasion by Trypanosoma cruzi Extracellular Amastigotes"

    Article Title: A Carbohydrate Moiety of Secreted Stage-Specific Glycoprotein 4 Participates in Host Cell Invasion by Trypanosoma cruzi Extracellular Amastigotes

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2018.00693

    Extracellular amastigotes release Ssp-4 associated with vesicles interacting with host cell. (A) Scanning electron microscopy of extracellular amastigotes (EAs) adhered to coverslips coated with poly-L-lysine or Vero cells. Scale bars: 2 and 5 μm. (B) Transmission electron microscopy of HeLa cells incubated for 30 min with EAs from the G strain (colored in red). Scale bar: 1 μm. Black arrows indicate EA secreted vesicles. (C) Immunofluorescence of HeLa cells incubated for 30 min with EAs from G (MOI 10:1) and mAb1D9 (green), DAPI (blue), and phalloidin-TRITC (red). Upper panels: images of a focal plane showing vesicles secreted by EAs of the G strain (green) associated with actin (red); bottom panels: three-dimensional reconstruction from a Z -series acquired by a confocal microscope. Scale bar: 2 μm. Arrows indicate vesicular structures secreted by EAs. (D) Extracellular amastigotes (EAs) from the G and Y strains were incubated for 6 h in RPMI medium without fetal bovine serum. Supernatant was collected and fractionated in three different populations: V2 (pelleted after 2 h of ultracentrifugation), V16 (pelleted after 16 h ultracentrifugation), and VF (supernatant from pellet V16). Western blotting with mAb1D9 and silver-stained SDS–PAGE of the different fractions. Whole cell (WC) lysate of EAs of the G strain was used as control.
    Figure Legend Snippet: Extracellular amastigotes release Ssp-4 associated with vesicles interacting with host cell. (A) Scanning electron microscopy of extracellular amastigotes (EAs) adhered to coverslips coated with poly-L-lysine or Vero cells. Scale bars: 2 and 5 μm. (B) Transmission electron microscopy of HeLa cells incubated for 30 min with EAs from the G strain (colored in red). Scale bar: 1 μm. Black arrows indicate EA secreted vesicles. (C) Immunofluorescence of HeLa cells incubated for 30 min with EAs from G (MOI 10:1) and mAb1D9 (green), DAPI (blue), and phalloidin-TRITC (red). Upper panels: images of a focal plane showing vesicles secreted by EAs of the G strain (green) associated with actin (red); bottom panels: three-dimensional reconstruction from a Z -series acquired by a confocal microscope. Scale bar: 2 μm. Arrows indicate vesicular structures secreted by EAs. (D) Extracellular amastigotes (EAs) from the G and Y strains were incubated for 6 h in RPMI medium without fetal bovine serum. Supernatant was collected and fractionated in three different populations: V2 (pelleted after 2 h of ultracentrifugation), V16 (pelleted after 16 h ultracentrifugation), and VF (supernatant from pellet V16). Western blotting with mAb1D9 and silver-stained SDS–PAGE of the different fractions. Whole cell (WC) lysate of EAs of the G strain was used as control.

    Techniques Used: Electron Microscopy, Transmission Assay, Incubation, Immunofluorescence, Microscopy, Western Blot, Staining, SDS Page

    16) Product Images from "A plasmid-based reporter system for live cell imaging of dengue infected cells"

    Article Title: A plasmid-based reporter system for live cell imaging of dengue infected cells

    Journal: Journal of virological methods

    doi: 10.1016/j.jviromet.2014.10.010

    Kinetics of nuclear localization of GFP after DENV infection. Vero cells transfected with p4B5-EGFP were infected with DENV-2 16681 and sequential images were acquired to assess the time for GFP to localize into the nucleus. Each set of images shows the expression of GFP in p4B5-EGFP transfected cells (top row), nuclear stain using NucBlue (middle row) and transmitted images (bottom row) at each time point. Time after addition of virus to the culture is located at the lower left of each image (magnification, ×20). Nuclear/total fluorescence (N/T) was calculated at each time point for a representative cell (arrow). The arrowhead shows a cell that remains uninfected over time. Data are representative of at least five experiments.
    Figure Legend Snippet: Kinetics of nuclear localization of GFP after DENV infection. Vero cells transfected with p4B5-EGFP were infected with DENV-2 16681 and sequential images were acquired to assess the time for GFP to localize into the nucleus. Each set of images shows the expression of GFP in p4B5-EGFP transfected cells (top row), nuclear stain using NucBlue (middle row) and transmitted images (bottom row) at each time point. Time after addition of virus to the culture is located at the lower left of each image (magnification, ×20). Nuclear/total fluorescence (N/T) was calculated at each time point for a representative cell (arrow). The arrowhead shows a cell that remains uninfected over time. Data are representative of at least five experiments.

    Techniques Used: Infection, Transfection, Expressing, Staining, Fluorescence

    Nuclear localization of GFP correlates with DENV antigen and co-expression of the NS2B3 protease. (a) Vero cells were transfected with p4B5-EGFP (green) and infected with DENV-2 16681 at an m.o.i of 1. 24 hours post-infection, cells were fixed, permeabilized and stained with antibody against DENV complex. The NS4B5-EGFP panel shows cytoplasmic expression of GFP (right, arrow neighboring cytoplasmic and nuclear expression of GFP (left, arrowhead) (magnification, ×100). DENV Ag panel shows DENV antigen staining in cells infected with DENV. Overlay of GFP and antigen staining (red) shows that nuclear localization of GFP correlates with DENV antigen staining (magnification, ×100). (b and c) Vero cells transfected with the p4B5-EGFP alone (b) or cotransfected with pNS2B3 (c) were analyzed for nuclear localization of GFP at 48hrs post-transfection (magnification, ×100). NS4B5-EGFP panel shows location of GFP within the cells (green). The nucleus panel shows the nucleus stained with NucBlue (blue, Life Technologies). The NS2B3 panel shows indirect antibody staining for NS3 (red). The overlay shows that nuclear GFP expression correlates with NS2B3 expression. Data are representative of at least six (a) and two (b) experiments.
    Figure Legend Snippet: Nuclear localization of GFP correlates with DENV antigen and co-expression of the NS2B3 protease. (a) Vero cells were transfected with p4B5-EGFP (green) and infected with DENV-2 16681 at an m.o.i of 1. 24 hours post-infection, cells were fixed, permeabilized and stained with antibody against DENV complex. The NS4B5-EGFP panel shows cytoplasmic expression of GFP (right, arrow neighboring cytoplasmic and nuclear expression of GFP (left, arrowhead) (magnification, ×100). DENV Ag panel shows DENV antigen staining in cells infected with DENV. Overlay of GFP and antigen staining (red) shows that nuclear localization of GFP correlates with DENV antigen staining (magnification, ×100). (b and c) Vero cells transfected with the p4B5-EGFP alone (b) or cotransfected with pNS2B3 (c) were analyzed for nuclear localization of GFP at 48hrs post-transfection (magnification, ×100). NS4B5-EGFP panel shows location of GFP within the cells (green). The nucleus panel shows the nucleus stained with NucBlue (blue, Life Technologies). The NS2B3 panel shows indirect antibody staining for NS3 (red). The overlay shows that nuclear GFP expression correlates with NS2B3 expression. Data are representative of at least six (a) and two (b) experiments.

    Techniques Used: Expressing, Transfection, Infection, Staining

    All four DENV serotypes induce cleavage of p4B5-EGFP to localize GFP to the nucleus. Vero cells transfected with p4B5-EGFP (green) were infected with each of the four DENV serotypes at an MOI of 1. Cells were fixed, permeabilized and stained for DENV antigen (red) and nuclear DNA (cyan). Cells were analyzed at 24 hours post-infection by fluorescence microscopy. Each row is a representative image of DENV infected cells in bright field (trans) and fluorescence images showing nuclear stain (cyan), DENV antigen stain (red), 4B5-EGFP expression (green). The overlay is a composite of the nucleus, DENV and NS4B-EGFP images. (a) DENV-1 Hawaii, (b) DENV-2 C0112/96, (c) DENV-3 CH53489 and (d) DENV-4 814669. Data are representative of at least four experiments each.
    Figure Legend Snippet: All four DENV serotypes induce cleavage of p4B5-EGFP to localize GFP to the nucleus. Vero cells transfected with p4B5-EGFP (green) were infected with each of the four DENV serotypes at an MOI of 1. Cells were fixed, permeabilized and stained for DENV antigen (red) and nuclear DNA (cyan). Cells were analyzed at 24 hours post-infection by fluorescence microscopy. Each row is a representative image of DENV infected cells in bright field (trans) and fluorescence images showing nuclear stain (cyan), DENV antigen stain (red), 4B5-EGFP expression (green). The overlay is a composite of the nucleus, DENV and NS4B-EGFP images. (a) DENV-1 Hawaii, (b) DENV-2 C0112/96, (c) DENV-3 CH53489 and (d) DENV-4 814669. Data are representative of at least four experiments each.

    Techniques Used: Transfection, Infection, Staining, Fluorescence, Microscopy, Expressing

    Construction and characterization of p4B5-EGFP (a) Schematic of the p4B5-EGFP construct containing NS4B and the first 30 nucleotides of NS5 of the DENV-2 genome tagged with the SV40 NLS and eGFP. The arrow indicates the cleavage site utilized by the DENV NS2B3 protease. The vertical lines represent restriction endonuclease sites. (b) Unfixed Vero cells transfected with p4B5-EGFP were stained with ER-Tracker Blue-White DPX dye (Cyan) 24hrs post-transfection and immediately imaged to detect colocalization of NS4B5-EGFP (reporter, green) with ER membranes (blue, magnification, ×100). Inset reflects ER staining of cells alone. (c) Unfixed Vero cells transfected with p4B5-EGFP were infected with DENV-2 16681. Nuclei were counterstained with NucBlue for live cells (Invitrogen) (blue) to detect nuclear localization of GFP (green, magnification, ×100). Cytoplasmic expression of GFP is indicated by the arrowhead and nuclear expression of GFP is indicated by an arrow (magnification, ×100). (d) Analysis of nuclear to total fluorescence intensity ratios of GFP (nuclear:total fl) in uninfected and DENV infected cells at 24 h was performed using ImageJ software. Each symbol represents analysis of a single cell. Statistical analysis was performed using nonparametric Wilcoxon-Mann-Whitney rank sum test. Straight line represents the median. (e) Cellular lysates were prepared 24 h post infection and subjected to Western blotting for GFP or β-actin. The 5-EGFP fragment represents the cleavage product resulting from cleavage between NS4B and NS5-EGFP during viral infection.
    Figure Legend Snippet: Construction and characterization of p4B5-EGFP (a) Schematic of the p4B5-EGFP construct containing NS4B and the first 30 nucleotides of NS5 of the DENV-2 genome tagged with the SV40 NLS and eGFP. The arrow indicates the cleavage site utilized by the DENV NS2B3 protease. The vertical lines represent restriction endonuclease sites. (b) Unfixed Vero cells transfected with p4B5-EGFP were stained with ER-Tracker Blue-White DPX dye (Cyan) 24hrs post-transfection and immediately imaged to detect colocalization of NS4B5-EGFP (reporter, green) with ER membranes (blue, magnification, ×100). Inset reflects ER staining of cells alone. (c) Unfixed Vero cells transfected with p4B5-EGFP were infected with DENV-2 16681. Nuclei were counterstained with NucBlue for live cells (Invitrogen) (blue) to detect nuclear localization of GFP (green, magnification, ×100). Cytoplasmic expression of GFP is indicated by the arrowhead and nuclear expression of GFP is indicated by an arrow (magnification, ×100). (d) Analysis of nuclear to total fluorescence intensity ratios of GFP (nuclear:total fl) in uninfected and DENV infected cells at 24 h was performed using ImageJ software. Each symbol represents analysis of a single cell. Statistical analysis was performed using nonparametric Wilcoxon-Mann-Whitney rank sum test. Straight line represents the median. (e) Cellular lysates were prepared 24 h post infection and subjected to Western blotting for GFP or β-actin. The 5-EGFP fragment represents the cleavage product resulting from cleavage between NS4B and NS5-EGFP during viral infection.

    Techniques Used: Construct, Transfection, Staining, Infection, Expressing, Fluorescence, Software, MANN-WHITNEY, Western Blot

    17) Product Images from "Highly potent anti-HIV-1 activity isolated from fermented Polygonum tinctorium Aiton"

    Article Title: Highly potent anti-HIV-1 activity isolated from fermented Polygonum tinctorium Aiton

    Journal: Antiviral Research

    doi: 10.1016/j.antiviral.2005.02.003

    Anti-viral activity of Sukumo extract. (A) Anti-HIV-1 activity of Sukumo extract in MT-4 cells was measured by MTT assay. HIV-1 (III B ) was used in this study ((●) mock infected and (♦) HIV-1)) and the EC 50 values for inhibition of Sukumo extract against HIV-1 replication were determined. (B) Anti-HSV-1 activity of Sukumo extract in Vero cells was determined by plaque assay. The results shown are mean ± S.D. of triplicates.
    Figure Legend Snippet: Anti-viral activity of Sukumo extract. (A) Anti-HIV-1 activity of Sukumo extract in MT-4 cells was measured by MTT assay. HIV-1 (III B ) was used in this study ((●) mock infected and (♦) HIV-1)) and the EC 50 values for inhibition of Sukumo extract against HIV-1 replication were determined. (B) Anti-HSV-1 activity of Sukumo extract in Vero cells was determined by plaque assay. The results shown are mean ± S.D. of triplicates.

    Techniques Used: Activity Assay, MTT Assay, Infection, Inhibition, Plaque Assay

    18) Product Images from "A Novel Mode of Poxvirus Superinfection Exclusion That Prevents Fusion of the Lipid Bilayers of Viral and Cellular Membranes"

    Article Title: A Novel Mode of Poxvirus Superinfection Exclusion That Prevents Fusion of the Lipid Bilayers of Viral and Cellular Membranes

    Journal: Journal of Virology

    doi: 10.1128/JVI.00816-14

    Selectivity of SIE. (A) CPXV. Cells were uninfected or infected with 10 PFU/cell of VACV strain WR or IHD-J or CPXV strain Brighton at 37°C for various lengths of time and then superinfected with 3 PFU/cell of WRvFire, IHD-JvFire, or CPXVvFire at 37°C for 150 min. LUC activity then was measured and plotted as a percentage of the uninfected cell control. (B) WNV. Cells were uninfected or infected with VACV at 10 PFU/cell of IHD-J at 37°C for 180 min. WNV reporter virus particles (3 particles per cell) were added, and cells were incubated at 37°C for 20 h. Cells were lysed and renilla LUC activity in cellular extracts measured. (C) VSV. Equivalent numbers of HeLa and Vero cells were uninfected or infected with 10 PFU/cell VACV IHD-J for 240 min at 37°C. Cells then were superinfected with 3 PFU/cell of VSV for 21 h at 37°C. Cells were lysed, and VSV G and cellular GAPDH proteins were analyzed by Western blotting. As a specificity control, cells were treated with dynasore to prevent VSV entry and G protein expression. Image J was used to quantify VSV G protein in each sample as arbitrary units.
    Figure Legend Snippet: Selectivity of SIE. (A) CPXV. Cells were uninfected or infected with 10 PFU/cell of VACV strain WR or IHD-J or CPXV strain Brighton at 37°C for various lengths of time and then superinfected with 3 PFU/cell of WRvFire, IHD-JvFire, or CPXVvFire at 37°C for 150 min. LUC activity then was measured and plotted as a percentage of the uninfected cell control. (B) WNV. Cells were uninfected or infected with VACV at 10 PFU/cell of IHD-J at 37°C for 180 min. WNV reporter virus particles (3 particles per cell) were added, and cells were incubated at 37°C for 20 h. Cells were lysed and renilla LUC activity in cellular extracts measured. (C) VSV. Equivalent numbers of HeLa and Vero cells were uninfected or infected with 10 PFU/cell VACV IHD-J for 240 min at 37°C. Cells then were superinfected with 3 PFU/cell of VSV for 21 h at 37°C. Cells were lysed, and VSV G and cellular GAPDH proteins were analyzed by Western blotting. As a specificity control, cells were treated with dynasore to prevent VSV entry and G protein expression. Image J was used to quantify VSV G protein in each sample as arbitrary units.

    Techniques Used: Infection, Activity Assay, Incubation, Western Blot, Expressing

    19) Product Images from "The potential molecular effects of bursal septpeptide II on immune induction and antitumor activity"

    Article Title: The potential molecular effects of bursal septpeptide II on immune induction and antitumor activity

    Journal: Journal of Veterinary Science

    doi: 10.4142/jvs.2015.16.3.325

    p53 transcription and protein expression after BSP-II treatment. Vero cells were transfected with p53 Luc and pRL-TK plasmid, and then cultured with or without BSP-II for 24 h. p53-luciferase activity levels were then measured (A). The transfected Vero cells were also pre-incubated with α-pifithrin for 2 h, incubated with or without BSP-II (2 µg/mL) for 22 h, and the level of p53-luciferase activity was measured (B). Non-transfected Vero cells were cultured with or without BSP-II for 24 h, and Western blotting analysis was performed to detect p53 and Bax protein expressions (C). Transfected or non-transfected Vero cells were treated with Dox as a positive control. Bars represent the mean ± SD of three independent experiments. * p
    Figure Legend Snippet: p53 transcription and protein expression after BSP-II treatment. Vero cells were transfected with p53 Luc and pRL-TK plasmid, and then cultured with or without BSP-II for 24 h. p53-luciferase activity levels were then measured (A). The transfected Vero cells were also pre-incubated with α-pifithrin for 2 h, incubated with or without BSP-II (2 µg/mL) for 22 h, and the level of p53-luciferase activity was measured (B). Non-transfected Vero cells were cultured with or without BSP-II for 24 h, and Western blotting analysis was performed to detect p53 and Bax protein expressions (C). Transfected or non-transfected Vero cells were treated with Dox as a positive control. Bars represent the mean ± SD of three independent experiments. * p

    Techniques Used: Expressing, Transfection, Plasmid Preparation, Cell Culture, Luciferase, Activity Assay, Incubation, Western Blot, Positive Control

    20) Product Images from "TMPRSS2 and MSPL Facilitate Trypsin-Independent Porcine Epidemic Diarrhea Virus Replication in Vero Cells"

    Article Title: TMPRSS2 and MSPL Facilitate Trypsin-Independent Porcine Epidemic Diarrhea Virus Replication in Vero Cells

    Journal: Viruses

    doi: 10.3390/v9050114

    Effects of TTSPs and TTSP inhibitor on viral replication. ( A ) Porcine epidemic diarrhea virus (PEDV) titers following the expression of TTSPs in Vero cells. Viral titers were determined by plaque assay. *** p
    Figure Legend Snippet: Effects of TTSPs and TTSP inhibitor on viral replication. ( A ) Porcine epidemic diarrhea virus (PEDV) titers following the expression of TTSPs in Vero cells. Viral titers were determined by plaque assay. *** p

    Techniques Used: Expressing, Plaque Assay

    Expression of type II transmembrane serine proteases (TTSPs) in transfected Vero cells. ( A ) Post-transfection, the expression of TMPRSS2, HAT, DESC1, and MSPL in transfected Vero cells was detected via indirect immunofluorescence. Bar = 25 μm. Magnification, ×200; ( B ) TTSP expression in transfected Vero cells as determined by western blot. Zymogens and the mature form are indicated; ( C ) TTSPs expression was detected by FACS. The geometric mean channel fluorescence (GMCF) measured in a representative experiment performed with triplicate samples is shown. Error bars indicate standard deviations of three independent experiments.
    Figure Legend Snippet: Expression of type II transmembrane serine proteases (TTSPs) in transfected Vero cells. ( A ) Post-transfection, the expression of TMPRSS2, HAT, DESC1, and MSPL in transfected Vero cells was detected via indirect immunofluorescence. Bar = 25 μm. Magnification, ×200; ( B ) TTSP expression in transfected Vero cells as determined by western blot. Zymogens and the mature form are indicated; ( C ) TTSPs expression was detected by FACS. The geometric mean channel fluorescence (GMCF) measured in a representative experiment performed with triplicate samples is shown. Error bars indicate standard deviations of three independent experiments.

    Techniques Used: Expressing, Transfection, HAT Assay, Immunofluorescence, Western Blot, FACS, Fluorescence

    21) Product Images from "Caspase-Mediated Cleavage of Nucleocapsid protein of a Protease-Independent Porcine Epidemic Diarrhea Virus Strain"

    Article Title: Caspase-Mediated Cleavage of Nucleocapsid protein of a Protease-Independent Porcine Epidemic Diarrhea Virus Strain

    Journal: Virus Research

    doi: 10.1016/j.virusres.2020.198026

    Subcellular localization of PEDV N protein in Vero cells. Vero cells were mock-infected or infected at MOI of 1 with PEDV 8aa with DMSO, 8aa with Z-VAD-fmk (100 μM) or KD with trypsin (1 µg/ml). For the expression of N protein, 24 h-old semi-confluent cells were transfected with pIRES-N-nHA, and cells were incubated at 37 °C for (A) 12 h (KD) or 36 h (8aa and N transfection), or (B)12, 24 or 48 h (8aa and 8aa with Z-VAD-fmk). Fixed and permeabilized cells were stained with anti-PEDV N monoclonal antibody (green) and SYTOX orange (Blue) and examined by confocal microscopy.
    Figure Legend Snippet: Subcellular localization of PEDV N protein in Vero cells. Vero cells were mock-infected or infected at MOI of 1 with PEDV 8aa with DMSO, 8aa with Z-VAD-fmk (100 μM) or KD with trypsin (1 µg/ml). For the expression of N protein, 24 h-old semi-confluent cells were transfected with pIRES-N-nHA, and cells were incubated at 37 °C for (A) 12 h (KD) or 36 h (8aa and N transfection), or (B)12, 24 or 48 h (8aa and 8aa with Z-VAD-fmk). Fixed and permeabilized cells were stained with anti-PEDV N monoclonal antibody (green) and SYTOX orange (Blue) and examined by confocal microscopy.

    Techniques Used: Infection, Expressing, Transfection, Incubation, Staining, Confocal Microscopy

    22) Product Images from "African Swine Fever Virus IAP Homologue Inhibits Caspase Activation and Promotes Cell Survival in Mammalian Cells"

    Article Title: African Swine Fever Virus IAP Homologue Inhibits Caspase Activation and Promotes Cell Survival in Mammalian Cells

    Journal: Journal of Virology

    doi: 10.1128/JVI.75.6.2535-2543.2001

    ASFV IAP protects Vero cells from TNF-CHX or staurosporine-induced apoptosis. Cells were stably transfected with control pcDNA vector or vector containing the A224L gene. (A) DEVD cleavage activity was measured using extracts from transfected Vero cells incubated or not with TNF-α (100 ng/ml) and CHX (5 μg/ml) for 16 h. The results are the average of three experiments (error bars, standard deviation). (B) Transfected Vero cells were incubated with medium or medium plus 1 μM staurosporine for 24 h, and the DNA content of the cells was determined by flow cytometry after propidium iodide staining.
    Figure Legend Snippet: ASFV IAP protects Vero cells from TNF-CHX or staurosporine-induced apoptosis. Cells were stably transfected with control pcDNA vector or vector containing the A224L gene. (A) DEVD cleavage activity was measured using extracts from transfected Vero cells incubated or not with TNF-α (100 ng/ml) and CHX (5 μg/ml) for 16 h. The results are the average of three experiments (error bars, standard deviation). (B) Transfected Vero cells were incubated with medium or medium plus 1 μM staurosporine for 24 h, and the DNA content of the cells was determined by flow cytometry after propidium iodide staining.

    Techniques Used: Stable Transfection, Transfection, Plasmid Preparation, Activity Assay, Incubation, Standard Deviation, Flow Cytometry, Cytometry, Staining

    23) Product Images from "Ras-GAP Binding and Phosphorylation by Herpes Simplex Virus Type 2 RR1 PK (ICP10) and Activation of the Ras/MEK/MAPK Mitogenic Pathway Are Required for Timely Onset of Virus Growth"

    Article Title: Ras-GAP Binding and Phosphorylation by Herpes Simplex Virus Type 2 RR1 PK (ICP10) and Activation of the Ras/MEK/MAPK Mitogenic Pathway Are Required for Timely Onset of Virus Growth

    Journal: Journal of Virology

    doi:

    Onset of HSV-2 growth is delayed by PD98059. Vero cells were pretreated (1 h, 37°C) with 50 μM PD98059 (○) or PD98059 reconstitution medium (●) and infected with HSV-2. They were reincubated in medium containing 1% FCS with or without PD98059, respectively. Adsorption was for 1 h at 37°C (0 h of growth curve). Virus titers were determined at 2 to 48 h after adsorption, and the results are expressed as the burst size (PFU/cell ± the standard error of the mean). Cells infected with ICP10ΔPK in medium containing 10% FCS (▵) were studied in parallel.
    Figure Legend Snippet: Onset of HSV-2 growth is delayed by PD98059. Vero cells were pretreated (1 h, 37°C) with 50 μM PD98059 (○) or PD98059 reconstitution medium (●) and infected with HSV-2. They were reincubated in medium containing 1% FCS with or without PD98059, respectively. Adsorption was for 1 h at 37°C (0 h of growth curve). Virus titers were determined at 2 to 48 h after adsorption, and the results are expressed as the burst size (PFU/cell ± the standard error of the mean). Cells infected with ICP10ΔPK in medium containing 10% FCS (▵) were studied in parallel.

    Techniques Used: Infection, Adsorption

    24) Product Images from "Excoecarianin, Isolated from Phyllanthus urinaria Linnea, Inhibits Herpes Simplex Virus Type 2 Infection through Inactivation of Viral Particles"

    Article Title: Excoecarianin, Isolated from Phyllanthus urinaria Linnea, Inhibits Herpes Simplex Virus Type 2 Infection through Inactivation of Viral Particles

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    doi: 10.1093/ecam/nep157

    The morphology of Vero cells without treatment (a) and with treatment of ACV (b) or excoecarianin (c)-(d). Vero cells were seeded onto 24-well culture plates at density of 1 × 10 3 cells per well. After 4 h, excoecarianin or ACV was added. The cells were incubated for 7 days, and the cellular morphology was examined under phase-contrast microscope.
    Figure Legend Snippet: The morphology of Vero cells without treatment (a) and with treatment of ACV (b) or excoecarianin (c)-(d). Vero cells were seeded onto 24-well culture plates at density of 1 × 10 3 cells per well. After 4 h, excoecarianin or ACV was added. The cells were incubated for 7 days, and the cellular morphology was examined under phase-contrast microscope.

    Techniques Used: Incubation, Microscopy

    Inhibitory effect of excoecarianin (open bars) and ACV (dotted bars) against HSV-2 infection in Vero cell as determined by plaque reduction assay. Vero cells were incubated with 100 pfu of HSV-2 and different concentrations of excoecarianin or ACV. After 1 h, an overlay medium containing 1% methylcellulose was added. On Day 3 post-infection, the cell monolayer was stained with crystal violet and the virus plaques formed were counted. The percentage of inhibition was calculated by comparing the plaque number of compound-treated group to that of the untreated group. The concentrations of excoecarianin and ACV that inhibited 50% of HSV-2 infection (IC 50 ) were determined. Each bar represents the mean ± SD of three independent experiments. The asterisk indicates significant difference between test sample and solvent control ( P
    Figure Legend Snippet: Inhibitory effect of excoecarianin (open bars) and ACV (dotted bars) against HSV-2 infection in Vero cell as determined by plaque reduction assay. Vero cells were incubated with 100 pfu of HSV-2 and different concentrations of excoecarianin or ACV. After 1 h, an overlay medium containing 1% methylcellulose was added. On Day 3 post-infection, the cell monolayer was stained with crystal violet and the virus plaques formed were counted. The percentage of inhibition was calculated by comparing the plaque number of compound-treated group to that of the untreated group. The concentrations of excoecarianin and ACV that inhibited 50% of HSV-2 infection (IC 50 ) were determined. Each bar represents the mean ± SD of three independent experiments. The asterisk indicates significant difference between test sample and solvent control ( P

    Techniques Used: Infection, Incubation, Staining, Inhibition

    Synergistic antiviral activity of ACV and excoecarianin in Vero cells. IC 50 values were derived from the data shown in Table 3 and used to construct the isobologram. FIC 50 of ACV represents the ratio of the IC 50 of ACV in the presence of a constant concentration of excoecarianin to the IC 50 of ACV alone. The x -axis represents the ratio of the fixed concentration of excoecarianin to the IC 50 of excoecarianin alone. In this representation, displacement of the experimental data points to the left of the theoretical line is indicative of synergistic behavior.
    Figure Legend Snippet: Synergistic antiviral activity of ACV and excoecarianin in Vero cells. IC 50 values were derived from the data shown in Table 3 and used to construct the isobologram. FIC 50 of ACV represents the ratio of the IC 50 of ACV in the presence of a constant concentration of excoecarianin to the IC 50 of ACV alone. The x -axis represents the ratio of the fixed concentration of excoecarianin to the IC 50 of excoecarianin alone. In this representation, displacement of the experimental data points to the left of the theoretical line is indicative of synergistic behavior.

    Techniques Used: Activity Assay, Derivative Assay, Construct, Concentration Assay

    The cytotoxic effect of excoecarianin (open bars) and ACV (dotted bars) toward Vero cells as determined by XTT assay. Various concentrations of excoecarianin or ACV were added to Vero cells. After 72 h of incubation, the XTT solution was added and then the optical densities were measured. The cytotoxic effect of excoecarianin and ACV were evaluated and the 50% cytotoxic concentration (CC 50 ) was calculated. Each bar represents the mean ± SD of three independent experiments. The asterisk indicates significant difference between test sample and solvent control ( P
    Figure Legend Snippet: The cytotoxic effect of excoecarianin (open bars) and ACV (dotted bars) toward Vero cells as determined by XTT assay. Various concentrations of excoecarianin or ACV were added to Vero cells. After 72 h of incubation, the XTT solution was added and then the optical densities were measured. The cytotoxic effect of excoecarianin and ACV were evaluated and the 50% cytotoxic concentration (CC 50 ) was calculated. Each bar represents the mean ± SD of three independent experiments. The asterisk indicates significant difference between test sample and solvent control ( P

    Techniques Used: XTT Assay, Incubation, Concentration Assay

    25) Product Images from "Nonmuscle Myosin Heavy Chain IIA Is a Critical Factor Contributing to the Efficiency of Early Infection of Severe Fever with Thrombocytopenia Syndrome Virus"

    Article Title: Nonmuscle Myosin Heavy Chain IIA Is a Critical Factor Contributing to the Efficiency of Early Infection of Severe Fever with Thrombocytopenia Syndrome Virus

    Journal: Journal of Virology

    doi: 10.1128/JVI.02141-13

    Interaction of NMMHC-IIA with SFTSV glycoprotein Gn. (A) Vero cells were lysed in PBS containing 1.0% β-DDM and incubated with Dynal protein A Sepharose beads in the presence of 15 μg of purified Gn-mFc or CD147-mFc fusion protein. Bound
    Figure Legend Snippet: Interaction of NMMHC-IIA with SFTSV glycoprotein Gn. (A) Vero cells were lysed in PBS containing 1.0% β-DDM and incubated with Dynal protein A Sepharose beads in the presence of 15 μg of purified Gn-mFc or CD147-mFc fusion protein. Bound

    Techniques Used: Incubation, Purification

    26) Product Images from "Preventing Cleavage of the Respiratory Syncytial Virus Attachment Protein in Vero Cells Rescues the Infectivity of Progeny Virus for Primary Human Airway Cultures"

    Article Title: Preventing Cleavage of the Respiratory Syncytial Virus Attachment Protein in Vero Cells Rescues the Infectivity of Progeny Virus for Primary Human Airway Cultures

    Journal: Journal of Virology

    doi: 10.1128/JVI.02351-15

    Cathepsin L and B expression and activity in HeLa and Vero cells. (A) Cathepsin L mRNA from mock-treated or rgRSV-infected HeLa and Vero cells was reverse transcribed and amplified by reverse transcription-PCR (RT-PCR), and the specific 294-bp product
    Figure Legend Snippet: Cathepsin L and B expression and activity in HeLa and Vero cells. (A) Cathepsin L mRNA from mock-treated or rgRSV-infected HeLa and Vero cells was reverse transcribed and amplified by reverse transcription-PCR (RT-PCR), and the specific 294-bp product

    Techniques Used: Expressing, Activity Assay, Infection, Amplification, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction

    Cathepsin L treatment of purified virions that had been grown in the presence of a cathepsin L inhibitor (inh). Vero cells (A) or HeLa cells (B) were inoculated with rgRSV and treated with medium containing vehicle (first and third lanes) or 0.5 μM
    Figure Legend Snippet: Cathepsin L treatment of purified virions that had been grown in the presence of a cathepsin L inhibitor (inh). Vero cells (A) or HeLa cells (B) were inoculated with rgRSV and treated with medium containing vehicle (first and third lanes) or 0.5 μM

    Techniques Used: Purification

    Protease inhibition of G-protein cleavage in Vero cells. Immunoblotting was used to detect biotinylated cell surface RSV G protein (55 or 90 kDa) produced in rgRSV-inoculated Vero cells treated with the following protease inhibitors: 2-fold dilutions
    Figure Legend Snippet: Protease inhibition of G-protein cleavage in Vero cells. Immunoblotting was used to detect biotinylated cell surface RSV G protein (55 or 90 kDa) produced in rgRSV-inoculated Vero cells treated with the following protease inhibitors: 2-fold dilutions

    Techniques Used: Inhibition, Produced

    Comparison of rgRSV and rgRSV-L208A infection of HAE cells. HeLa cell-derived (H-D) and Vero cell-derived (V-D) rgRSV and rgRSV-L208A were produced in HeLa or Vero cells. (A) Virus was purified by use of a sucrose gradient, and G protein was detected
    Figure Legend Snippet: Comparison of rgRSV and rgRSV-L208A infection of HAE cells. HeLa cell-derived (H-D) and Vero cell-derived (V-D) rgRSV and rgRSV-L208A were produced in HeLa or Vero cells. (A) Virus was purified by use of a sucrose gradient, and G protein was detected

    Techniques Used: Infection, Derivative Assay, Produced, Purification

    Ability of rgRSV grown in the presence of a cathepsin L inhibitor to infect HAE cells. The infectivity for HAE cells of partially purified rgRSV virions that were produced in HeLa or Vero cells in the presence of a cathepsin L inhibitor or DMSO (
    Figure Legend Snippet: Ability of rgRSV grown in the presence of a cathepsin L inhibitor to infect HAE cells. The infectivity for HAE cells of partially purified rgRSV virions that were produced in HeLa or Vero cells in the presence of a cathepsin L inhibitor or DMSO (

    Techniques Used: Infection, Purification, Produced

    27) Product Images from "Replication of Herpes Simplex Virus: Egress of Progeny Virus at Specialized Cell Membrane Sites"

    Article Title: Replication of Herpes Simplex Virus: Egress of Progeny Virus at Specialized Cell Membrane Sites

    Journal: Journal of Virology

    doi: 10.1128/JVI.00463-12

    Glycoprotein enrichment at coverslip-adherent surface egress sites. (A) TIRF micrographs of a VP26-GFP HSV-1-infected Vero cell at 12 hpi fixed, permeabilized, and stained with rhodamine-conjugated WGA (Rh-WGA) to mark glycosylated proteins. As in EM pictures, GFP-labeled virions were found to cluster at specific sites along the cell surface. Note that glycosylated proteins accumulate at these sites. A magnified image of the box is shown in panel E. (B) TIRF micrographs of a VP26-GFP HSV-infected Vero cell at 12 hpi that had been treated with α-gB (DL16) antibody. (C) TIRF micrographs of a VP26-GFP HSV-infected Vero cell treated with α-gD antibody (DL11). Staining indicates that viral glycoproteins accumulate at egress sites. Arrowheads mark two egress sites where the pocket-like structure of the sites is apparent. The holes in the donuts are areas where the membrane has extended beyond the 300-nm laser excitation range. Virus is rarely visible in the holes, indicating that virions are closely bound to the cell membrane. (D) Mock-infected Vero cell permeabilized and stained with WGA shown at that same exposure as that for the infected cell in panel A. The cell edge is outlined as determined by increasing the brightness of the image until the edge was visible. The size bar in panel C is also relevant for panels A to D. (F) HSV-1-infected Vero cell stained with both rhodamine-WGA and α-gD antibody. Note that there is complete colocalization between the two. Alexa 594-conjugated secondary antibodies were used in panels B, C, and F.
    Figure Legend Snippet: Glycoprotein enrichment at coverslip-adherent surface egress sites. (A) TIRF micrographs of a VP26-GFP HSV-1-infected Vero cell at 12 hpi fixed, permeabilized, and stained with rhodamine-conjugated WGA (Rh-WGA) to mark glycosylated proteins. As in EM pictures, GFP-labeled virions were found to cluster at specific sites along the cell surface. Note that glycosylated proteins accumulate at these sites. A magnified image of the box is shown in panel E. (B) TIRF micrographs of a VP26-GFP HSV-infected Vero cell at 12 hpi that had been treated with α-gB (DL16) antibody. (C) TIRF micrographs of a VP26-GFP HSV-infected Vero cell treated with α-gD antibody (DL11). Staining indicates that viral glycoproteins accumulate at egress sites. Arrowheads mark two egress sites where the pocket-like structure of the sites is apparent. The holes in the donuts are areas where the membrane has extended beyond the 300-nm laser excitation range. Virus is rarely visible in the holes, indicating that virions are closely bound to the cell membrane. (D) Mock-infected Vero cell permeabilized and stained with WGA shown at that same exposure as that for the infected cell in panel A. The cell edge is outlined as determined by increasing the brightness of the image until the edge was visible. The size bar in panel C is also relevant for panels A to D. (F) HSV-1-infected Vero cell stained with both rhodamine-WGA and α-gD antibody. Note that there is complete colocalization between the two. Alexa 594-conjugated secondary antibodies were used in panels B, C, and F.

    Techniques Used: Infection, Staining, Whole Genome Amplification, Labeling

    TIRF micrographs showing formation of glycoprotein patch sites uncoupled from virus egress. (A) Vero cells infected with VP26-GFP HSV-1 were fixed at 4, 6, and 8 hpi. Patch glycoproteins were labeled with rhodamine-WGA. All images were taken at the same exposure. In the 4-hpi image, the edge of the adherent surface of the cell (as determined by increasing the brightness of the image) is outlined. Note that neither glycoprotein patches nor virus are visible at 4 hpi. By 6 hpi, glycoprotein patches are beginning to form (arrow) on the adherent surface, but virus still is not evident. At 8 hpi, there is pronounced glycoprotein staining on the cell surface and virions are beginning to accumulate in these areas. Images indicate that patches form before viral egress. (B) Vero cells infected with wild-type KOS HSV-1 (top) or UL25Δ mutant HSV (bottom) were fixed at 12 hpi. In a UL25Δ infection, capsids are retained in the nucleus. Egress site glycoproteins were labeled with WGA. Virus was labeled with α-VP5 major capsid protein antibody and Alexa 488-labeled secondary antibody. Infections suggest that adherent surface glycoprotein patches form independently of viral release.
    Figure Legend Snippet: TIRF micrographs showing formation of glycoprotein patch sites uncoupled from virus egress. (A) Vero cells infected with VP26-GFP HSV-1 were fixed at 4, 6, and 8 hpi. Patch glycoproteins were labeled with rhodamine-WGA. All images were taken at the same exposure. In the 4-hpi image, the edge of the adherent surface of the cell (as determined by increasing the brightness of the image) is outlined. Note that neither glycoprotein patches nor virus are visible at 4 hpi. By 6 hpi, glycoprotein patches are beginning to form (arrow) on the adherent surface, but virus still is not evident. At 8 hpi, there is pronounced glycoprotein staining on the cell surface and virions are beginning to accumulate in these areas. Images indicate that patches form before viral egress. (B) Vero cells infected with wild-type KOS HSV-1 (top) or UL25Δ mutant HSV (bottom) were fixed at 12 hpi. In a UL25Δ infection, capsids are retained in the nucleus. Egress site glycoproteins were labeled with WGA. Virus was labeled with α-VP5 major capsid protein antibody and Alexa 488-labeled secondary antibody. Infections suggest that adherent surface glycoprotein patches form independently of viral release.

    Techniques Used: Infection, Labeling, Whole Genome Amplification, Staining, Mutagenesis

    Location of progeny virions in HSV-1-infected Vero cells. (A to D) Thin-section electron micrographs of infected Vero cells fixed and processed on coverslips at 12 hpi. Note that the majority of virions are released in pockets along the adherent cell surface (arrowheads) and at cell-cell contact points (arrows), with some virus above cell-cell contacts (bracket). Panel B is an enlargement of the left arrowhead in panel A; panel C is an enlargement of the right arrowhead in panel A. (E and F) Electron micrographs of mock-infected Vero cells. Note that, unlike infected cells, the cell membrane is closely apposed to the coverslip edge. In panels A to F the coverslip interface is marked by the thin line near the bottom surface of the cell. (G) Consecutive Z-stack confocal images of a representative glass coverslip-grown Vero cell infected with VP26-GFP mutant virus. Sections begin at the coverslip and go up by 0.25-μm increments. Asterisks mark the two sections above the coverslip where the majority of virions can be seen. Other sections have far fewer virions. The nucleus (containing VP26-GFP-labeled capsid assembly areas) is outlined. Arrows indicate virus released along a cell-cell contact. Images were obtained with a Philips 400T transmission electron microscope (A to F) and a Zeiss LSM 510 confocal microscope with an inverted 100× lens (G).
    Figure Legend Snippet: Location of progeny virions in HSV-1-infected Vero cells. (A to D) Thin-section electron micrographs of infected Vero cells fixed and processed on coverslips at 12 hpi. Note that the majority of virions are released in pockets along the adherent cell surface (arrowheads) and at cell-cell contact points (arrows), with some virus above cell-cell contacts (bracket). Panel B is an enlargement of the left arrowhead in panel A; panel C is an enlargement of the right arrowhead in panel A. (E and F) Electron micrographs of mock-infected Vero cells. Note that, unlike infected cells, the cell membrane is closely apposed to the coverslip edge. In panels A to F the coverslip interface is marked by the thin line near the bottom surface of the cell. (G) Consecutive Z-stack confocal images of a representative glass coverslip-grown Vero cell infected with VP26-GFP mutant virus. Sections begin at the coverslip and go up by 0.25-μm increments. Asterisks mark the two sections above the coverslip where the majority of virions can be seen. Other sections have far fewer virions. The nucleus (containing VP26-GFP-labeled capsid assembly areas) is outlined. Arrows indicate virus released along a cell-cell contact. Images were obtained with a Philips 400T transmission electron microscope (A to F) and a Zeiss LSM 510 confocal microscope with an inverted 100× lens (G).

    Techniques Used: Infection, Mutagenesis, Labeling, Transmission Assay, Microscopy

    Effect of gE and gE cytoplasmic tail deletions on patch formation and progeny virion trafficking. (A) TIRF images of Vero cells infected with the ΔgE mutant, the gEΔCT mutant, or the gEΔCT rescue virus at an MOI of 0.3 for 12 h. Cells were fixed and treated with rhodamine-WGA and α-VP5 antibody with Alexa 488-conjugated secondary antibody. Note that patches are smaller in size in the gE deletion mutant-infected cells, but the virion number in those patches is increased at 12 hpi. (B) Quantitative determination of the percentage of adherent cell membrane covered by glycoprotein patches and (C) the amount of virions on the total adherent membrane. Quantifications support the visual data. (D) Infected cells were harvested at 10 hpi, and the titer was determined by limiting-dilution plaque assay. Note that gE deletions had little effect on the production of progeny virus titer despite the observation that the number of virions on the adherent surface was increased. P values of deletion mutants compared to rescue virus are labeled the following: *,
    Figure Legend Snippet: Effect of gE and gE cytoplasmic tail deletions on patch formation and progeny virion trafficking. (A) TIRF images of Vero cells infected with the ΔgE mutant, the gEΔCT mutant, or the gEΔCT rescue virus at an MOI of 0.3 for 12 h. Cells were fixed and treated with rhodamine-WGA and α-VP5 antibody with Alexa 488-conjugated secondary antibody. Note that patches are smaller in size in the gE deletion mutant-infected cells, but the virion number in those patches is increased at 12 hpi. (B) Quantitative determination of the percentage of adherent cell membrane covered by glycoprotein patches and (C) the amount of virions on the total adherent membrane. Quantifications support the visual data. (D) Infected cells were harvested at 10 hpi, and the titer was determined by limiting-dilution plaque assay. Note that gE deletions had little effect on the production of progeny virus titer despite the observation that the number of virions on the adherent surface was increased. P values of deletion mutants compared to rescue virus are labeled the following: *,

    Techniques Used: Infection, Mutagenesis, Whole Genome Amplification, Plaque Assay, Labeling

    TIRF micrographs showing egress site reformation after actin depolymerization-induced disruption. VP26-GFP-infected Vero cells at 11.5 hpi (A) were treated with 1.7 μg/ml cytochalasin B for 30 min (B). (C) Toxin was then rinsed out and the infection continued for another 45 min. Sample cells were fixed at each step and stained with Texas Red-phalloidin or rhodamine-WGA. (D) Untreated control. Note that both the actin cortex and viral egress sites were able to reorganize after disruption. There is no global block in actin polymerization.
    Figure Legend Snippet: TIRF micrographs showing egress site reformation after actin depolymerization-induced disruption. VP26-GFP-infected Vero cells at 11.5 hpi (A) were treated with 1.7 μg/ml cytochalasin B for 30 min (B). (C) Toxin was then rinsed out and the infection continued for another 45 min. Sample cells were fixed at each step and stained with Texas Red-phalloidin or rhodamine-WGA. (D) Untreated control. Note that both the actin cortex and viral egress sites were able to reorganize after disruption. There is no global block in actin polymerization.

    Techniques Used: Infection, Staining, Whole Genome Amplification, Blocking Assay

    28) Product Images from "Nucleocytoplasmic Shuttling of the HSV-2 Serine/Threonine Kinase Us3"

    Article Title: Nucleocytoplasmic Shuttling of the HSV-2 Serine/Threonine Kinase Us3

    Journal: Virology

    doi: 10.1016/j.virol.2011.06.011

    Analysis of nuclear export of HSV-2 Us3 by fluorescence loss in photobleaching assays (FLIP). Vero cells were transfected with plasmids encoding either mCherry or a HSV-2 Us3 mCherry fusion protein (Us3-mCherry). At 48 hours post transfection, images
    Figure Legend Snippet: Analysis of nuclear export of HSV-2 Us3 by fluorescence loss in photobleaching assays (FLIP). Vero cells were transfected with plasmids encoding either mCherry or a HSV-2 Us3 mCherry fusion protein (Us3-mCherry). At 48 hours post transfection, images

    Techniques Used: Fluorescence, Transfection

    29) Product Images from "Characterization of Glycoprotein-Mediated Entry of Severe Fever with Thrombocytopenia Syndrome Virus"

    Article Title: Characterization of Glycoprotein-Mediated Entry of Severe Fever with Thrombocytopenia Syndrome Virus

    Journal: Journal of Virology

    doi: 10.1128/JVI.00110-16

    Inhibition of SFTSVpv and SFTSV infection by H + -ATPase inhibitors. Huh7 or Vero cells were inoculated with SFTSVpv, RVFVpv, VSVpv, or MLVpv (A) or with SFTSV (B) after treatment of these cells with various concentrations of ammonium chloride (left) or
    Figure Legend Snippet: Inhibition of SFTSVpv and SFTSV infection by H + -ATPase inhibitors. Huh7 or Vero cells were inoculated with SFTSVpv, RVFVpv, VSVpv, or MLVpv (A) or with SFTSV (B) after treatment of these cells with various concentrations of ammonium chloride (left) or

    Techniques Used: Inhibition, Infection

    Expression and localization of SFTSV-GP. (A) Expression of SFTSV-GP in Vero (top) or Huh7 (bottom) cells infected with SFTSV (left) or transfected with a GP plasmid (right) was examined by an immunofluorescence assay with an anti-SFTSV-GP monoclonal antibody.
    Figure Legend Snippet: Expression and localization of SFTSV-GP. (A) Expression of SFTSV-GP in Vero (top) or Huh7 (bottom) cells infected with SFTSV (left) or transfected with a GP plasmid (right) was examined by an immunofluorescence assay with an anti-SFTSV-GP monoclonal antibody.

    Techniques Used: Expressing, Infection, Transfection, Plasmid Preparation, Immunofluorescence

    Effects of 25HC on infection by SFTSVpv and SFTSV. (A) Inhibition of SFTSVpv infection by 25HC. Vero (left) and Huh7 (right) cells were inoculated with pseudotype viruses after treatment of these cells with various concentrations of 25HC. Infectivities
    Figure Legend Snippet: Effects of 25HC on infection by SFTSVpv and SFTSV. (A) Inhibition of SFTSVpv infection by 25HC. Vero (left) and Huh7 (right) cells were inoculated with pseudotype viruses after treatment of these cells with various concentrations of 25HC. Infectivities

    Techniques Used: Infection, Inhibition

    30) Product Images from "Temporal Transcriptome and Promoter Architecture of the African Swine Fever Virus"

    Article Title: Temporal Transcriptome and Promoter Architecture of the African Swine Fever Virus

    Journal: bioRxiv

    doi: 10.1101/847343

    Early and late gene expression of ASFV BA71V. (a) FPKM values for 20 most highly expressed ASFV TUs according to CAGE-seq at 5h (left) and 16h (right) post-infection, RNA-seq equivalent results found in SUPPLEMENTARY Figure 3. Genes highlighted in maroon indicate those encoding proteins which were also found in the 20 most-abundantly expressed AFSV proteins during infection of either WSL-HP, HEK293 or Vero cells according to proteome analysis done by Keßler et al . 31 . Gene functions are shown after their name with TR and PSP referring to predicted transmembrane region and putative signal peptide, respectively. (b) MAplot from DESeq2 analysis of CAGE-seq representing the DESeq2 base mean of transcript levels versus their log2 fold change, with significantly differentially expressed genes in purple (adjusted p -value
    Figure Legend Snippet: Early and late gene expression of ASFV BA71V. (a) FPKM values for 20 most highly expressed ASFV TUs according to CAGE-seq at 5h (left) and 16h (right) post-infection, RNA-seq equivalent results found in SUPPLEMENTARY Figure 3. Genes highlighted in maroon indicate those encoding proteins which were also found in the 20 most-abundantly expressed AFSV proteins during infection of either WSL-HP, HEK293 or Vero cells according to proteome analysis done by Keßler et al . 31 . Gene functions are shown after their name with TR and PSP referring to predicted transmembrane region and putative signal peptide, respectively. (b) MAplot from DESeq2 analysis of CAGE-seq representing the DESeq2 base mean of transcript levels versus their log2 fold change, with significantly differentially expressed genes in purple (adjusted p -value

    Techniques Used: Expressing, Infection, RNA Sequencing Assay

    31) Product Images from "TMPRSS2 and MSPL Facilitate Trypsin-Independent Porcine Epidemic Diarrhea Virus Replication in Vero Cells"

    Article Title: TMPRSS2 and MSPL Facilitate Trypsin-Independent Porcine Epidemic Diarrhea Virus Replication in Vero Cells

    Journal: Viruses

    doi: 10.3390/v9050114

    Effects of TTSPs and TTSP inhibitor on viral replication. ( A ) Porcine epidemic diarrhea virus (PEDV) titers following the expression of TTSPs in Vero cells. Viral titers were determined by plaque assay. *** p
    Figure Legend Snippet: Effects of TTSPs and TTSP inhibitor on viral replication. ( A ) Porcine epidemic diarrhea virus (PEDV) titers following the expression of TTSPs in Vero cells. Viral titers were determined by plaque assay. *** p

    Techniques Used: Expressing, Plaque Assay

    Culture of PEDV isolated from pig intestine in Vero cells transiently expressing TTSPs in three serial passages. ( A ) Isolation of PEDV strain A; ( B ) Isolation of PEDV strain B. The relative quantity of pCMV-Myc without trypsin at the 1st passage = 1. Error bars indicate standard error of the mean. Results shown are those of a representative experiment performed with triplicate samples.
    Figure Legend Snippet: Culture of PEDV isolated from pig intestine in Vero cells transiently expressing TTSPs in three serial passages. ( A ) Isolation of PEDV strain A; ( B ) Isolation of PEDV strain B. The relative quantity of pCMV-Myc without trypsin at the 1st passage = 1. Error bars indicate standard error of the mean. Results shown are those of a representative experiment performed with triplicate samples.

    Techniques Used: Isolation, Expressing

    Expression of type II transmembrane serine proteases (TTSPs) in transfected Vero cells. ( A ) Post-transfection, the expression of TMPRSS2, HAT, DESC1, and MSPL in transfected Vero cells was detected via indirect immunofluorescence. Bar = 25 μm. Magnification, ×200; ( B ) TTSP expression in transfected Vero cells as determined by western blot. Zymogens and the mature form are indicated; ( C ) TTSPs expression was detected by FACS. The geometric mean channel fluorescence (GMCF) measured in a representative experiment performed with triplicate samples is shown. Error bars indicate standard deviations of three independent experiments.
    Figure Legend Snippet: Expression of type II transmembrane serine proteases (TTSPs) in transfected Vero cells. ( A ) Post-transfection, the expression of TMPRSS2, HAT, DESC1, and MSPL in transfected Vero cells was detected via indirect immunofluorescence. Bar = 25 μm. Magnification, ×200; ( B ) TTSP expression in transfected Vero cells as determined by western blot. Zymogens and the mature form are indicated; ( C ) TTSPs expression was detected by FACS. The geometric mean channel fluorescence (GMCF) measured in a representative experiment performed with triplicate samples is shown. Error bars indicate standard deviations of three independent experiments.

    Techniques Used: Expressing, Transfection, HAT Assay, Immunofluorescence, Western Blot, FACS, Fluorescence

    32) Product Images from "Chemical Induction of Endogenous Retrovirus Particles from the Vero Cell Line of African Green Monkeys ▿"

    Article Title: Chemical Induction of Endogenous Retrovirus Particles from the Vero Cell Line of African Green Monkeys ▿

    Journal: Journal of Virology

    doi: 10.1128/JVI.00147-11

    Drug dose evaluation and PERT activity. Drug dose range for IUdR (A), AzaC (B), and NaBut (C) was determined by evaluating the Vero cell viability after drug treatment with various concentrations for 48 h. Cell toxicity was determined at day 0 (day of
    Figure Legend Snippet: Drug dose evaluation and PERT activity. Drug dose range for IUdR (A), AzaC (B), and NaBut (C) was determined by evaluating the Vero cell viability after drug treatment with various concentrations for 48 h. Cell toxicity was determined at day 0 (day of

    Techniques Used: Activity Assay

    Infectivity analysis of drug-induced retrovirus from Vero cells. Vero cells (1 × 10 6 ) were planted in 25-cm 2 flasks for 16 h and treated with 1.25 μg/ml AzaC or 200 μg/ml IUdR for 48 h. In the case of one flask, medium was replaced
    Figure Legend Snippet: Infectivity analysis of drug-induced retrovirus from Vero cells. Vero cells (1 × 10 6 ) were planted in 25-cm 2 flasks for 16 h and treated with 1.25 μg/ml AzaC or 200 μg/ml IUdR for 48 h. In the case of one flask, medium was replaced

    Techniques Used: Infection

    33) Product Images from "Trypanocidal Activity of Smallanthus sonchifolius: Identification of Active Sesquiterpene Lactones by Bioassay-Guided Fractionation"

    Article Title: Trypanocidal Activity of Smallanthus sonchifolius: Identification of Active Sesquiterpene Lactones by Bioassay-Guided Fractionation

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    doi: 10.1155/2013/627898

    Cytotoxicity of enhydrin, uvedalin, and polymatin B on Vero cells. Cultures were kept for 24 h in the presence of different concentrations (1 to 50 μ g/mL) of the STLs. Cell viability was determined by the MTT method and was expressed as the ratio between viable cells in the presence and absence of the compound multiplied by 100. Bars represent the mean ± SEM of three experiments carried out in duplicate.
    Figure Legend Snippet: Cytotoxicity of enhydrin, uvedalin, and polymatin B on Vero cells. Cultures were kept for 24 h in the presence of different concentrations (1 to 50 μ g/mL) of the STLs. Cell viability was determined by the MTT method and was expressed as the ratio between viable cells in the presence and absence of the compound multiplied by 100. Bars represent the mean ± SEM of three experiments carried out in duplicate.

    Techniques Used: MTT Assay

    34) Product Images from "Inflammatory monocytes mediate control of acute alphavirus infection in mice"

    Article Title: Inflammatory monocytes mediate control of acute alphavirus infection in mice

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1006748

    Induction of type I IFN expression in monocytes requires virus infection. (A-B) Enriched WT bone marrow monocytes were co-cultured with Vero cells infected with RRV-GFP (n = 3/group). After 18 h of co-culture, GFP expression in Vero cells and monocytes was measured by flow cytometry. (A) Shown are representative histograms. (B) The percent GFP + monocytes and Vero cells. Data are representative of two independent experiments. (C-D) Enriched WT bone marrow monocytes were co-cultured with uninfected or RRV-GFP-infected Vero cells in the presence or absence of 1 μM Latrunculin B. After 18 h of co-culture, (C) IFNα2 mRNA expression level in monocytes was quantified by qRT-PCR. Data are normalized to 18S rRNA levels and are expressed as the relative expression ( n -fold increase) over expression in uninfected Vero cells without monocytes. (D) The percent GFP + Vero cells and monocytes were measured by flow cytometry based on GFP expression within the CD45 - Vero cells and the CD45 + monocytes. Data are combined from two independent experiments. P values were determined by one-way ANOVA with a Tukey’s multiple comparison test.
    Figure Legend Snippet: Induction of type I IFN expression in monocytes requires virus infection. (A-B) Enriched WT bone marrow monocytes were co-cultured with Vero cells infected with RRV-GFP (n = 3/group). After 18 h of co-culture, GFP expression in Vero cells and monocytes was measured by flow cytometry. (A) Shown are representative histograms. (B) The percent GFP + monocytes and Vero cells. Data are representative of two independent experiments. (C-D) Enriched WT bone marrow monocytes were co-cultured with uninfected or RRV-GFP-infected Vero cells in the presence or absence of 1 μM Latrunculin B. After 18 h of co-culture, (C) IFNα2 mRNA expression level in monocytes was quantified by qRT-PCR. Data are normalized to 18S rRNA levels and are expressed as the relative expression ( n -fold increase) over expression in uninfected Vero cells without monocytes. (D) The percent GFP + Vero cells and monocytes were measured by flow cytometry based on GFP expression within the CD45 - Vero cells and the CD45 + monocytes. Data are combined from two independent experiments. P values were determined by one-way ANOVA with a Tukey’s multiple comparison test.

    Techniques Used: Expressing, Infection, Cell Culture, Co-Culture Assay, Flow Cytometry, Cytometry, Quantitative RT-PCR, Over Expression

    35) Product Images from "Adaptation of High-Growth Influenza H5N1 Vaccine Virus in Vero Cells: Implications for Pandemic Preparedness"

    Article Title: Adaptation of High-Growth Influenza H5N1 Vaccine Virus in Vero Cells: Implications for Pandemic Preparedness

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0024057

    Plaque morphology of NIBRG-14 and Vero cell-adapted (Vero-15) H5N1 viruses grown in Vero cells in different days post infection (DPI).
    Figure Legend Snippet: Plaque morphology of NIBRG-14 and Vero cell-adapted (Vero-15) H5N1 viruses grown in Vero cells in different days post infection (DPI).

    Techniques Used: Infection

    36) Product Images from "Zaire Ebola virus entry into human dendritic cells is insensitive to cathepsin L inhibition"

    Article Title: Zaire Ebola virus entry into human dendritic cells is insensitive to cathepsin L inhibition

    Journal: Cellular microbiology

    doi: 10.1111/j.1462-5822.2009.01385.x

    Effect of cathepsin B and L inhibitors on Ebola VLP entry and EBOV infection of VEROs and DCs VERO (A, C, E) and DCs (B, D) were pretreated with mock and decreasing concentrations of cathepsin B and cathepsin L, respectively. Also, VERO (A) and DCs (B) were mock and VLP treated or EBOV GFP -infected (denoted by bar under graphs). Solid bars represent relative percentage of VEROs (A) and DCs (B) infected by VLPs and white bars represent relative percentage of VEROs (A) and DCs (B) infected by EBOV GFP . Cathepsin B (C, D) and cathepsin L (E) activity was measured in duplicate (solid and white bars) using a fluorogenic substrate and expressed as relative fluorenscence units (RFU) using equivalent amounts of pretreatred VEROs (C, E) and DCs (D). (F) Equivalent amounts of VERO (lane 1) and DC (lane 2) lysates (same lysates used to determine cathepsin activity) were subjected to SDS-PAGE and western blotting using anti-cathepsin L antibody (204106, R D systems, Minneapolis, MN). Arrow denotes positions of Cathepsin L species. (G) VP40+GP VLPs were incubated for 1.5 hours with decreasing concentrations of recombinant cathepsin L (5, 1, 0.2, 0.04 μg/mL) and subjected to western blotting using a polyclonal anti-GP antibody. Arrow denotes positions of GP species.
    Figure Legend Snippet: Effect of cathepsin B and L inhibitors on Ebola VLP entry and EBOV infection of VEROs and DCs VERO (A, C, E) and DCs (B, D) were pretreated with mock and decreasing concentrations of cathepsin B and cathepsin L, respectively. Also, VERO (A) and DCs (B) were mock and VLP treated or EBOV GFP -infected (denoted by bar under graphs). Solid bars represent relative percentage of VEROs (A) and DCs (B) infected by VLPs and white bars represent relative percentage of VEROs (A) and DCs (B) infected by EBOV GFP . Cathepsin B (C, D) and cathepsin L (E) activity was measured in duplicate (solid and white bars) using a fluorogenic substrate and expressed as relative fluorenscence units (RFU) using equivalent amounts of pretreatred VEROs (C, E) and DCs (D). (F) Equivalent amounts of VERO (lane 1) and DC (lane 2) lysates (same lysates used to determine cathepsin activity) were subjected to SDS-PAGE and western blotting using anti-cathepsin L antibody (204106, R D systems, Minneapolis, MN). Arrow denotes positions of Cathepsin L species. (G) VP40+GP VLPs were incubated for 1.5 hours with decreasing concentrations of recombinant cathepsin L (5, 1, 0.2, 0.04 μg/mL) and subjected to western blotting using a polyclonal anti-GP antibody. Arrow denotes positions of GP species.

    Techniques Used: Infection, Activity Assay, SDS Page, Western Blot, Incubation, Recombinant

    37) Product Images from "A 176 amino acid polypeptide derived from the mumps virus HN ectodomain shows immunological and biological properties similar to the HN protein"

    Article Title: A 176 amino acid polypeptide derived from the mumps virus HN ectodomain shows immunological and biological properties similar to the HN protein

    Journal: Virology Journal

    doi: 10.1186/1743-422X-7-195

    Expression of the pcDNAHN176-construct . A) Detection of HN176 mRNA in transfected Vero cells by RT-PCR amplification. Lane 1) ϕX174 DNA- HaeIII marker; Lane 2) RT/PCR negative control; Lane 3) Positive control (MuV-infected cells); Lane 4) Vero-untransfected cells, 5) Vero cells transfected with the plasmid pcDNA3.1, Lane 6) Vero cells transfected with the pcDNA-HN176-construct. The arrow indicates 580 bp amplicon. 1% agarose gel/100 V/1 hr/49 mA. B ) RT-PCR Controls. Lane 1) ϕX174 DNA- HaeIII marker; Lane 2) PCR amplification of the RNA samples obtained from pcDNAHN176-transfected Vero cells without a previous RT reaction; Lane 3 and 4) RT/PCR amplification of β-actin gene using RNA samples obtained from HeLa cells and pcDNAHN176-transfected Vero cells, respectively. C ) Immunodetection of the HN176 polypeptide by immunochemistry (1 st row) and immunofluorescence (2 nd row). Frames 1 5 mock infected cells; 2 6 pcDNA3.1-transfected cells; 3 7 MuV-infected cells; 4 8 pcDNAHN176-transfected cells, the blue (immunochemistry) and the green (immunofluorescence) colors indicate a positive reaction, N indicates the nucleus. 40×
    Figure Legend Snippet: Expression of the pcDNAHN176-construct . A) Detection of HN176 mRNA in transfected Vero cells by RT-PCR amplification. Lane 1) ϕX174 DNA- HaeIII marker; Lane 2) RT/PCR negative control; Lane 3) Positive control (MuV-infected cells); Lane 4) Vero-untransfected cells, 5) Vero cells transfected with the plasmid pcDNA3.1, Lane 6) Vero cells transfected with the pcDNA-HN176-construct. The arrow indicates 580 bp amplicon. 1% agarose gel/100 V/1 hr/49 mA. B ) RT-PCR Controls. Lane 1) ϕX174 DNA- HaeIII marker; Lane 2) PCR amplification of the RNA samples obtained from pcDNAHN176-transfected Vero cells without a previous RT reaction; Lane 3 and 4) RT/PCR amplification of β-actin gene using RNA samples obtained from HeLa cells and pcDNAHN176-transfected Vero cells, respectively. C ) Immunodetection of the HN176 polypeptide by immunochemistry (1 st row) and immunofluorescence (2 nd row). Frames 1 5 mock infected cells; 2 6 pcDNA3.1-transfected cells; 3 7 MuV-infected cells; 4 8 pcDNAHN176-transfected cells, the blue (immunochemistry) and the green (immunofluorescence) colors indicate a positive reaction, N indicates the nucleus. 40×

    Techniques Used: Expressing, Construct, Transfection, Reverse Transcription Polymerase Chain Reaction, Amplification, Marker, Negative Control, Positive Control, Infection, Plasmid Preparation, Agarose Gel Electrophoresis, Polymerase Chain Reaction, Immunodetection, Immunofluorescence

    Hemadsorption and neuraminidase activities in the pcDNAHN176 transfected cells . A) . Hemadsorption (HD) reaction. a) Mock infected cells; b) MuV-infected cells; c) pcDNA-3.1-transfected cells; d) pcDNAHN176-transfected Vero cells. The red cell aggregates indicate a positive HD. 40×. B ) Neuraminidase (NA) reaction in total cellular protein extract by a dot blot assay: a) Mock infected cells; b) MuV-infected cells; c) pcDNA-3.1-transfected cells; d) pcDNAHN176-transfected Vero cells. C) % of hemoglobin (Hb) released after the HD. The Hb absorbance of MuV-infected cells was considered as 100% and was used to calculate the % of Hb for the different cells. D ) Comparison of the NA activity in MuV-infected cells, pcDNA3.1-transfected cells and pcDNAHN176-transfected Vero cells by dot blot and spectrophotometric methods.
    Figure Legend Snippet: Hemadsorption and neuraminidase activities in the pcDNAHN176 transfected cells . A) . Hemadsorption (HD) reaction. a) Mock infected cells; b) MuV-infected cells; c) pcDNA-3.1-transfected cells; d) pcDNAHN176-transfected Vero cells. The red cell aggregates indicate a positive HD. 40×. B ) Neuraminidase (NA) reaction in total cellular protein extract by a dot blot assay: a) Mock infected cells; b) MuV-infected cells; c) pcDNA-3.1-transfected cells; d) pcDNAHN176-transfected Vero cells. C) % of hemoglobin (Hb) released after the HD. The Hb absorbance of MuV-infected cells was considered as 100% and was used to calculate the % of Hb for the different cells. D ) Comparison of the NA activity in MuV-infected cells, pcDNA3.1-transfected cells and pcDNAHN176-transfected Vero cells by dot blot and spectrophotometric methods.

    Techniques Used: Transfection, Infection, Dot Blot, Activity Assay

    Immunological properties of the pcDNAHN176-construct . A) Detection of the HN176 polypeptide and HN protein by Western Blot. Lane 1) Negative control, uninfected cells and sera from pcDNAHN176-immunized rabbits; Lane 2) MuV-infected-Vero cells and anti-MuV serum; Lane 3) pcDNAHN176-transfected cells and anti-MuV serum; Lane 4) MuV-infected-Vero cells and sera from pcDNAHN176-immunized rabbits; Lane 5) pcDNAHN176-transfected cells and sera from pcDNAHN176-immunized rabbits. The upper arrow indicates in lane 2 and 4 the position of the complete viral HN protein, and the lower arrow indicates the position of the HN176 polypeptide in lanes 3 5. B) Body gain weight of hamsters immunized and challenged with MuV. Group A, animals without immunization and uninfected; Group B, viral control group (animals without immunization and challenged with MuV); Group C, animals immunized with pcDNA3.1 without challenge; Group D, animals immunized with pcDNAHN176-construct without challenge; Group E, animals immunized with pcDNA3.1 and challenged with MuV; Group F, animals immunized with pcDNAHN176 and challenged with MuV. C) Virus isolation from different organs of the hamsters groups. MuV was detected by HD, quantifying the amount of Hb. D) Detection of IL associated to Th1 response in the hamsters groups. E) Detection of IL associated to Th2 response in the hamsters groups. ILs were measured using the Luminex System (Invitrogen ® ). F) Lymphoproliferation index of spleen cells obtained from the hamsters groups. Cell proliferation was measured by MTT method.
    Figure Legend Snippet: Immunological properties of the pcDNAHN176-construct . A) Detection of the HN176 polypeptide and HN protein by Western Blot. Lane 1) Negative control, uninfected cells and sera from pcDNAHN176-immunized rabbits; Lane 2) MuV-infected-Vero cells and anti-MuV serum; Lane 3) pcDNAHN176-transfected cells and anti-MuV serum; Lane 4) MuV-infected-Vero cells and sera from pcDNAHN176-immunized rabbits; Lane 5) pcDNAHN176-transfected cells and sera from pcDNAHN176-immunized rabbits. The upper arrow indicates in lane 2 and 4 the position of the complete viral HN protein, and the lower arrow indicates the position of the HN176 polypeptide in lanes 3 5. B) Body gain weight of hamsters immunized and challenged with MuV. Group A, animals without immunization and uninfected; Group B, viral control group (animals without immunization and challenged with MuV); Group C, animals immunized with pcDNA3.1 without challenge; Group D, animals immunized with pcDNAHN176-construct without challenge; Group E, animals immunized with pcDNA3.1 and challenged with MuV; Group F, animals immunized with pcDNAHN176 and challenged with MuV. C) Virus isolation from different organs of the hamsters groups. MuV was detected by HD, quantifying the amount of Hb. D) Detection of IL associated to Th1 response in the hamsters groups. E) Detection of IL associated to Th2 response in the hamsters groups. ILs were measured using the Luminex System (Invitrogen ® ). F) Lymphoproliferation index of spleen cells obtained from the hamsters groups. Cell proliferation was measured by MTT method.

    Techniques Used: Construct, Western Blot, Negative Control, Infection, Transfection, Virus Isolation Assay, Luminex, MTT Assay

    38) Product Images from "A Novel Borna Disease Virus Vector System That Stably Expresses Foreign Proteins from an Intercistronic Noncoding Region ▿"

    Article Title: A Novel Borna Disease Virus Vector System That Stably Expresses Foreign Proteins from an Intercistronic Noncoding Region ▿

    Journal: Journal of Virology

    doi: 10.1128/JVI.05554-11

    The G-deficient rBDV establishes persistent infection in cultured cells. (A) Growth kinetics of ΔGLLP/M-GFP virus in Vero-BG and Vero cells. The cell-free virus was inoculated onto the cells at an MOI of 0.01. Infected cells were split every 3
    Figure Legend Snippet: The G-deficient rBDV establishes persistent infection in cultured cells. (A) Growth kinetics of ΔGLLP/M-GFP virus in Vero-BG and Vero cells. The cell-free virus was inoculated onto the cells at an MOI of 0.01. Infected cells were split every 3

    Techniques Used: Infection, Cell Culture

    39) Product Images from "Chikungunya Virus Nonstructural Protein 2 Inhibits Type I/II Interferon-Stimulated JAK-STAT Signaling ▿Chikungunya Virus Nonstructural Protein 2 Inhibits Type I/II Interferon-Stimulated JAK-STAT Signaling ▿ †"

    Article Title: Chikungunya Virus Nonstructural Protein 2 Inhibits Type I/II Interferon-Stimulated JAK-STAT Signaling ▿Chikungunya Virus Nonstructural Protein 2 Inhibits Type I/II Interferon-Stimulated JAK-STAT Signaling ▿ †

    Journal: Journal of Virology

    doi: 10.1128/JVI.00949-10

    Mutation of a conserved proline in nsP2 abolishes the inhibitory effect of CHIKV and SINV replicons on JAK-STAT signaling. (A) Schematic representation of the CHIKrep-pac2AEGFP and SINrepLuc replicons. nsP2 mutations P718S and P726S are indicated with asterisks; pac, puromycin acetyltransferase. (B) Partial amino acid alignment of alphavirus nsP2s. RRV, Ross River virus; VEEV, Venezuelan equine encephalitis virus. The conserved proline and amino acid numbers within nsP2 proteins are indicated. (C) pSTAT1 nuclear translocation upon IFN-β induction in SINrepGFP (wild type and mutant nsP2-P726S)-transfected Vero cells. Cells were immunostained with an anti-pSTAT1 antibody. Open arrowheads indicate replicon-positive cells lacking nuclear pSTAT1; solid arrowheads indicate replicon-positive cells with nuclear pSTAT1. (D) Nuclear translocation of phospho-STAT1 upon IFN-β induction in CHIKrep-pac2AEGFP (wild type and mutant nsP2-P718S)-transfected Vero cells. Cells were immunostained with an anti-pSTAT1 antibody.
    Figure Legend Snippet: Mutation of a conserved proline in nsP2 abolishes the inhibitory effect of CHIKV and SINV replicons on JAK-STAT signaling. (A) Schematic representation of the CHIKrep-pac2AEGFP and SINrepLuc replicons. nsP2 mutations P718S and P726S are indicated with asterisks; pac, puromycin acetyltransferase. (B) Partial amino acid alignment of alphavirus nsP2s. RRV, Ross River virus; VEEV, Venezuelan equine encephalitis virus. The conserved proline and amino acid numbers within nsP2 proteins are indicated. (C) pSTAT1 nuclear translocation upon IFN-β induction in SINrepGFP (wild type and mutant nsP2-P726S)-transfected Vero cells. Cells were immunostained with an anti-pSTAT1 antibody. Open arrowheads indicate replicon-positive cells lacking nuclear pSTAT1; solid arrowheads indicate replicon-positive cells with nuclear pSTAT1. (D) Nuclear translocation of phospho-STAT1 upon IFN-β induction in CHIKrep-pac2AEGFP (wild type and mutant nsP2-P718S)-transfected Vero cells. Cells were immunostained with an anti-pSTAT1 antibody.

    Techniques Used: Mutagenesis, Translocation Assay, Transfection

    Inhibition of IFN-β-induced STAT1 nuclear translocation by individual CHIKV nsPs. (A) Schematic representation of the pCMV-nsP1, -2, -3, and -4 expression plasmids and the CHIKrep-mCherry replicon, expressing mCherry. CMV, cytomegalovirus immediate-early promoter; 2A, foot-and-mouth disease virus 2A autoprotease. The bacteriophage SP6 and CHIKV 26S promoters are indicated. (B) pSTAT1 nuclear translocation upon IFN-β induction in Vero cells transfected with pCMV-nsP1, -2, -3, or -4. Cells were immunostained with an anti-pSTAT1 antibody. (C) pSTAT1 nuclear translocation upon IFN-β induction in CHIKrep-mCherry-transfected Vero cells. Open arrowheads indicate cells positive for nsP1, -2, -3, or -4- or for the CHIKV replicon that lack nuclear pSTAT1; solid arrowheads indicate nsP1- to nsP4-positive cells with nuclear pSTAT1.
    Figure Legend Snippet: Inhibition of IFN-β-induced STAT1 nuclear translocation by individual CHIKV nsPs. (A) Schematic representation of the pCMV-nsP1, -2, -3, and -4 expression plasmids and the CHIKrep-mCherry replicon, expressing mCherry. CMV, cytomegalovirus immediate-early promoter; 2A, foot-and-mouth disease virus 2A autoprotease. The bacteriophage SP6 and CHIKV 26S promoters are indicated. (B) pSTAT1 nuclear translocation upon IFN-β induction in Vero cells transfected with pCMV-nsP1, -2, -3, or -4. Cells were immunostained with an anti-pSTAT1 antibody. (C) pSTAT1 nuclear translocation upon IFN-β induction in CHIKrep-mCherry-transfected Vero cells. Open arrowheads indicate cells positive for nsP1, -2, -3, or -4- or for the CHIKV replicon that lack nuclear pSTAT1; solid arrowheads indicate nsP1- to nsP4-positive cells with nuclear pSTAT1.

    Techniques Used: Inhibition, Translocation Assay, Expressing, Transfection

    Inhibition of type I/II IFN signaling and ISG induction by CHIKV infection. (A and B) Vero cells were transfected with a pRL-TK plasmid expressing Rluc and either a type I IFN-responsive (ISRE) or a type II IFN-responsive (GAS) Fluc reporter plasmid. At 24 h p.t., cells were infected with CHIKV at an MOI of 5 PFU/ml. At 4, 8, and 12 h p.i., cells were treated with IFN-α at 1,000 IU/ml (A) or with IFN-γ at 100 ng/ml (B) for 6 h; then they were assayed for Fluc and Rluc activities. Activities in mock-infected (uninfected) cells with/without IFN induction were also measured. Fluc values were divided by Rluc readings to compensate for virus-induced downregulation of transcription/translation and were expressed relative to values for mock-infected, IFN-treated samples. Average values from triplicate samples are shown. Error bars represent standard deviations. (C and D) Vero cells, either healthy or infected with CHIKV for 4, 8, or 12 h, were incubated with 1,000 IU of IFN-α (C) or 100 ng of IFN-γ (D) per ml for 10 h. Real-time RT-PCR values for the IFN-stimulated gene OAS2 were normalized to those for the housekeeping gene RPL13A. OAS2 mRNA transcription levels were expressed relative to those of mock-infected, IFN-treated samples. Average values from duplicate samples are shown. Error bars represent standard deviations.
    Figure Legend Snippet: Inhibition of type I/II IFN signaling and ISG induction by CHIKV infection. (A and B) Vero cells were transfected with a pRL-TK plasmid expressing Rluc and either a type I IFN-responsive (ISRE) or a type II IFN-responsive (GAS) Fluc reporter plasmid. At 24 h p.t., cells were infected with CHIKV at an MOI of 5 PFU/ml. At 4, 8, and 12 h p.i., cells were treated with IFN-α at 1,000 IU/ml (A) or with IFN-γ at 100 ng/ml (B) for 6 h; then they were assayed for Fluc and Rluc activities. Activities in mock-infected (uninfected) cells with/without IFN induction were also measured. Fluc values were divided by Rluc readings to compensate for virus-induced downregulation of transcription/translation and were expressed relative to values for mock-infected, IFN-treated samples. Average values from triplicate samples are shown. Error bars represent standard deviations. (C and D) Vero cells, either healthy or infected with CHIKV for 4, 8, or 12 h, were incubated with 1,000 IU of IFN-α (C) or 100 ng of IFN-γ (D) per ml for 10 h. Real-time RT-PCR values for the IFN-stimulated gene OAS2 were normalized to those for the housekeeping gene RPL13A. OAS2 mRNA transcription levels were expressed relative to those of mock-infected, IFN-treated samples. Average values from duplicate samples are shown. Error bars represent standard deviations.

    Techniques Used: Inhibition, Infection, Transfection, Plasmid Preparation, Expressing, Incubation, Quantitative RT-PCR

    (A to C) CHIKV infection blocks STAT1/STAT2 nuclear translocation without depleting endogenous STAT1 levels. Vero cells were infected by CHIKV and were treated with IFN-α (A and B) or IFN-γ (C) for 30 min. Cells were fixed and stained with monoclonal antibodies specific for CHIKV envelope protein and STAT1 (A and C) or STAT2 (B). (C) Block in nuclear translocation of STAT1 in CHIKV infection in response to treatment with IFN-γ. Arrowheads indicate cells negatively infected with CHIKV but with nuclear STAT1/2. (D) CHIKV infection blocks STAT1 phosphorylation in Vero cells in response to IFN treatment. pSTAT1, STAT1, and tubulin were detected by Western blotting in CHIKV-infected or mock-infected Vero cells that were either left untreated or induced with type I or type II IFNs. Lane 1, protein size marker (in kilodaltons).
    Figure Legend Snippet: (A to C) CHIKV infection blocks STAT1/STAT2 nuclear translocation without depleting endogenous STAT1 levels. Vero cells were infected by CHIKV and were treated with IFN-α (A and B) or IFN-γ (C) for 30 min. Cells were fixed and stained with monoclonal antibodies specific for CHIKV envelope protein and STAT1 (A and C) or STAT2 (B). (C) Block in nuclear translocation of STAT1 in CHIKV infection in response to treatment with IFN-γ. Arrowheads indicate cells negatively infected with CHIKV but with nuclear STAT1/2. (D) CHIKV infection blocks STAT1 phosphorylation in Vero cells in response to IFN treatment. pSTAT1, STAT1, and tubulin were detected by Western blotting in CHIKV-infected or mock-infected Vero cells that were either left untreated or induced with type I or type II IFNs. Lane 1, protein size marker (in kilodaltons).

    Techniques Used: Infection, Translocation Assay, Staining, Blocking Assay, Western Blot, Marker

    A CHIKV replicon efficiently inhibits type I/II IFN-induced JAK-STAT signaling independently of host shutoff. (A) Schematic representation of CHIKrepEGFP, expressing EGFP. (B) pSTAT1 nuclear translocation in Vero cells upon induction with type I and type II IFNs. (C) A CHIKV replicon blocks pSTAT1 nuclear translocation upon type I/II IFN induction. Vero cells were immunostained with an anti-pSTAT1 antibody 24 h p.t. (D) CHIKV RNA replication, but not translational shutoff, blocks STAT1 nuclear translocation. Vero cells were transfected with CHIKrep-EGFP replicon RNA in the absence or presence of cycloheximide (Chx). Cells were induced for 30 min with IFN-β at 12 h p.t. and were stained with an anti-STAT1 antibody. Open arrowheads indicate CHIKV replicon-positive cells lacking nuclear STAT1.
    Figure Legend Snippet: A CHIKV replicon efficiently inhibits type I/II IFN-induced JAK-STAT signaling independently of host shutoff. (A) Schematic representation of CHIKrepEGFP, expressing EGFP. (B) pSTAT1 nuclear translocation in Vero cells upon induction with type I and type II IFNs. (C) A CHIKV replicon blocks pSTAT1 nuclear translocation upon type I/II IFN induction. Vero cells were immunostained with an anti-pSTAT1 antibody 24 h p.t. (D) CHIKV RNA replication, but not translational shutoff, blocks STAT1 nuclear translocation. Vero cells were transfected with CHIKrep-EGFP replicon RNA in the absence or presence of cycloheximide (Chx). Cells were induced for 30 min with IFN-β at 12 h p.t. and were stained with an anti-STAT1 antibody. Open arrowheads indicate CHIKV replicon-positive cells lacking nuclear STAT1.

    Techniques Used: Expressing, Translocation Assay, Transfection, Staining

    40) Product Images from "Native High-Density Lipoproteins (HDL) with Higher Paraoxonase Exerts a Potent Antiviral Effect against SARS-CoV-2 (COVID-19), While Glycated HDL Lost the Antiviral Activity"

    Article Title: Native High-Density Lipoproteins (HDL) with Higher Paraoxonase Exerts a Potent Antiviral Effect against SARS-CoV-2 (COVID-19), While Glycated HDL Lost the Antiviral Activity

    Journal: Antioxidants

    doi: 10.3390/antiox10020209

    Comparison of glycated HDL and native HDL. ( A ) Cell viability of Vero E6 cell after treatment with lipoproteins. MTT cytotoxicity assays were carried out using uninfected Vero E6 cells with increasing concentrations of each lipoprotein. ( B ) Cytopathic effect (CPE) reduction activity of lipoproteins against SARS-CoV-2.
    Figure Legend Snippet: Comparison of glycated HDL and native HDL. ( A ) Cell viability of Vero E6 cell after treatment with lipoproteins. MTT cytotoxicity assays were carried out using uninfected Vero E6 cells with increasing concentrations of each lipoprotein. ( B ) Cytopathic effect (CPE) reduction activity of lipoproteins against SARS-CoV-2.

    Techniques Used: MTT Assay, Activity Assay

    Related Articles

    Viability Assay:

    Article Title: Ribavirin efficiently suppresses porcine nidovirus replication
    Article Snippet: The anti-β-actin antibody and horseradish peroxidase (HRP)-conjugated secondary antibody were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). .. 2.2 Cell viability assay The cytotoxic effects of reagents on PAM-pCD163 and Vero cells were analyzed by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (Sigma) detecting cell viability. .. Briefly, PAM-pCD163 and Vero cells were grown at 1 × 104 cells/well in a 96-well tissue culture plate with ribavirin or MPA treatment for 24 h. After one day of incubation, 50 μl of MTT solution (1.1 mg/ml) was added to each well and the samples were incubated for an additional 4 h. The supernatant was then removed from each well, after which 150 μl of DMSO was added to dissolve the color formazan crystal produced from the MTT.

    MTT Assay:

    Article Title: Ribavirin efficiently suppresses porcine nidovirus replication
    Article Snippet: The anti-β-actin antibody and horseradish peroxidase (HRP)-conjugated secondary antibody were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). .. 2.2 Cell viability assay The cytotoxic effects of reagents on PAM-pCD163 and Vero cells were analyzed by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (Sigma) detecting cell viability. .. Briefly, PAM-pCD163 and Vero cells were grown at 1 × 104 cells/well in a 96-well tissue culture plate with ribavirin or MPA treatment for 24 h. After one day of incubation, 50 μl of MTT solution (1.1 mg/ml) was added to each well and the samples were incubated for an additional 4 h. The supernatant was then removed from each well, after which 150 μl of DMSO was added to dissolve the color formazan crystal produced from the MTT.

    Centrifugation:

    Article Title: Cell culture isolation and sequence analysis of genetically diverse US porcine epidemic diarrhea virus strains including a novel strain with a large deletion in the spike gene
    Article Snippet: All Vero cells were grown in Dulbecco Modified Eagle Medium (DMEM, Life Technologies, Carlsbad, CA) supplemented with antibiotics (100 units/mL of penicillin, 100 μg/mL of streptomycin, and 0.25 μg/mL of Fungizone® ) (Life Technologies), and 5% heat inactivated fetal bovine serum (Hyclone, Logan, UT). .. 2.4 Preparation of PEDV inoculum for Vero cells PEDV-positive intestinal or fecal samples were diluted 10-fold in Dulbecco's Phosphate buffered Saline without Mg2+ and Ca2+ [PBS (−), Sigma, St. Louis, MO), and were vortexed briefly followed by centrifugation at 10,000 × g for 3 min at 4 °C. ..

    Western Blot:

    Article Title: An HSV-based library screen identifies PP1α as a negative TRPV1 regulator with analgesic activity in models of pain
    Article Snippet: PCR products were resolved on a 1% agarose gel, visualized by ethidium bromide staining, and isolated with the Qiaquick gel extraction kit. .. Western blotting Vero cells were infected with vT-GFP vector (MOI = 3), and infected cell lysates were prepared by incubating the cells in 1× RIPA lysis buffer (Millipore, Billerica, MA) with protease inhibitors (cOmplete, Mini; Roche, Indianapolis, IN) for 30 minutes on ice. .. Lysates were boiled for 5 minutes and resolved on a sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel at 100 V. Proteins were transferred onto a polyvinylidene fluoride (PVDF) membrane (Millipore).

    Infection:

    Article Title: An HSV-based library screen identifies PP1α as a negative TRPV1 regulator with analgesic activity in models of pain
    Article Snippet: PCR products were resolved on a 1% agarose gel, visualized by ethidium bromide staining, and isolated with the Qiaquick gel extraction kit. .. Western blotting Vero cells were infected with vT-GFP vector (MOI = 3), and infected cell lysates were prepared by incubating the cells in 1× RIPA lysis buffer (Millipore, Billerica, MA) with protease inhibitors (cOmplete, Mini; Roche, Indianapolis, IN) for 30 minutes on ice. .. Lysates were boiled for 5 minutes and resolved on a sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel at 100 V. Proteins were transferred onto a polyvinylidene fluoride (PVDF) membrane (Millipore).

    Article Title: Targeting an Oncolytic Influenza A Virus to Tumor Tissue by Elastase
    Article Snippet: Viruses The ΔNS1-H1N1 A/New Caledonia virus contains HA and NA from A/New Caledonia/20/99 (H1N1), PB1 of A/Texas/1/77 (H3N2), and PA, PB2, NP, M, and NS are of PR8 origin A/Puerto Rico/8/34 and were adapted by serial passages to grow on Vero cells in presence of 5 μg/ml porcine pancreatic elastase (Sigma-Aldrich, St. Louis, MO, USA). .. Trypsin-dependent NS116 -GFP/AT or elastase-dependent NS1116 -GFP/AE viruses were propagated at 37°C in Vero cells infected with MOI of 0.01 in the presence of either 1 μg L-1-Tosylamide-2-phenylethyl chloromethyl ketone (TPCK)-treated trypsin (Sigma Aldrich) ml−1 or 10 μg human neutrophil elastase (Serva Electrophoresis, Heidelberg, Germany) ml−1 . .. Virus titer was determined by a TCID50 assay.

    Article Title: Mucin-Like Domain of Ebola Virus Glycoprotein Enhances Selective Oncolytic Actions against Brain Tumors
    Article Snippet: SDS-PAGE and Western blot analysis. .. Vero cells infected with VSV-EBOV or VSV-EBOVΔMLD were lysed with radioimmunoprecipitation assay buffer (Sigma), and the protein concentration was quantified using a bicinchoninic acid protein assay kit (Thermo Scientific Pierce). ..

    Plasmid Preparation:

    Article Title: An HSV-based library screen identifies PP1α as a negative TRPV1 regulator with analgesic activity in models of pain
    Article Snippet: PCR products were resolved on a 1% agarose gel, visualized by ethidium bromide staining, and isolated with the Qiaquick gel extraction kit. .. Western blotting Vero cells were infected with vT-GFP vector (MOI = 3), and infected cell lysates were prepared by incubating the cells in 1× RIPA lysis buffer (Millipore, Billerica, MA) with protease inhibitors (cOmplete, Mini; Roche, Indianapolis, IN) for 30 minutes on ice. .. Lysates were boiled for 5 minutes and resolved on a sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel at 100 V. Proteins were transferred onto a polyvinylidene fluoride (PVDF) membrane (Millipore).

    Lysis:

    Article Title: An HSV-based library screen identifies PP1α as a negative TRPV1 regulator with analgesic activity in models of pain
    Article Snippet: PCR products were resolved on a 1% agarose gel, visualized by ethidium bromide staining, and isolated with the Qiaquick gel extraction kit. .. Western blotting Vero cells were infected with vT-GFP vector (MOI = 3), and infected cell lysates were prepared by incubating the cells in 1× RIPA lysis buffer (Millipore, Billerica, MA) with protease inhibitors (cOmplete, Mini; Roche, Indianapolis, IN) for 30 minutes on ice. .. Lysates were boiled for 5 minutes and resolved on a sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel at 100 V. Proteins were transferred onto a polyvinylidene fluoride (PVDF) membrane (Millipore).

    Incubation:

    Article Title: Incorporation of CD55 into the Zika Viral Envelope Contributes to Its Stability against Human Complement
    Article Snippet: The ct-value obtained by the positive control (mAb 4G2 against ZIKV E protein) was set as “1.0” and the relative change of the ct-units is given as “fold increase”. .. Flow Cytometric Analysis For the detection of membrane-bound complement regulatory proteins (mCRPs), virus-producing cells were immune-stained as follow: Vero cells, primary Fibroblasts, and A549 cells (5 × 105 cells/tube) were harvested by detaching with Accutase (Sigma, Vienna Austria), washed twice with buffer containing 2% FBS, 0.2% EDTA in PBS, and incubated with 10 µg/mL anti-CD59, anti-CD55, anti-CD46 or isotope control Abs for 1 h at 4 °C. .. After washing once in 1 mL FACS-buffer, a secondary goat anti-rabbit IgG (H + L) labeled with Pacific Blue (ORIGIN) was added and incubated for 30 min at 4 °C.

    Radio Immunoprecipitation:

    Article Title: Mucin-Like Domain of Ebola Virus Glycoprotein Enhances Selective Oncolytic Actions against Brain Tumors
    Article Snippet: SDS-PAGE and Western blot analysis. .. Vero cells infected with VSV-EBOV or VSV-EBOVΔMLD were lysed with radioimmunoprecipitation assay buffer (Sigma), and the protein concentration was quantified using a bicinchoninic acid protein assay kit (Thermo Scientific Pierce). ..

    Protein Concentration:

    Article Title: Mucin-Like Domain of Ebola Virus Glycoprotein Enhances Selective Oncolytic Actions against Brain Tumors
    Article Snippet: SDS-PAGE and Western blot analysis. .. Vero cells infected with VSV-EBOV or VSV-EBOVΔMLD were lysed with radioimmunoprecipitation assay buffer (Sigma), and the protein concentration was quantified using a bicinchoninic acid protein assay kit (Thermo Scientific Pierce). ..

    Bicinchoninic Acid Protein Assay:

    Article Title: Mucin-Like Domain of Ebola Virus Glycoprotein Enhances Selective Oncolytic Actions against Brain Tumors
    Article Snippet: SDS-PAGE and Western blot analysis. .. Vero cells infected with VSV-EBOV or VSV-EBOVΔMLD were lysed with radioimmunoprecipitation assay buffer (Sigma), and the protein concentration was quantified using a bicinchoninic acid protein assay kit (Thermo Scientific Pierce). ..

    Flow Cytometry:

    Article Title: The Cell Cycle and Virus Infection
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    Expressing:

    Article Title: The Cell Cycle and Virus Infection
    Article Snippet: As can be seen from the figure, caspase 8 is redistributed to the nucleolus in cells expressing a viral protein (IBVNHis) from an expression plasmid. .. Flow Cytometry of Caspase 8 For flow cytometry of caspase 8 expression, Vero cells either treated with 100 mM Etoposide for 48 h for transfected for 24 h (either with pTriEx 1.1 [Novagen] or pTriEx IBVNHis) are harvested by detaching the cells with 2 mL PBS/EDTA/trypsin and transferring them into canonical tubes containing 8 mL 10% DMEM. ..

    Transfection:

    Article Title: The Cell Cycle and Virus Infection
    Article Snippet: As can be seen from the figure, caspase 8 is redistributed to the nucleolus in cells expressing a viral protein (IBVNHis) from an expression plasmid. .. Flow Cytometry of Caspase 8 For flow cytometry of caspase 8 expression, Vero cells either treated with 100 mM Etoposide for 48 h for transfected for 24 h (either with pTriEx 1.1 [Novagen] or pTriEx IBVNHis) are harvested by detaching the cells with 2 mL PBS/EDTA/trypsin and transferring them into canonical tubes containing 8 mL 10% DMEM. ..

    Transferring:

    Article Title: The Cell Cycle and Virus Infection
    Article Snippet: As can be seen from the figure, caspase 8 is redistributed to the nucleolus in cells expressing a viral protein (IBVNHis) from an expression plasmid. .. Flow Cytometry of Caspase 8 For flow cytometry of caspase 8 expression, Vero cells either treated with 100 mM Etoposide for 48 h for transfected for 24 h (either with pTriEx 1.1 [Novagen] or pTriEx IBVNHis) are harvested by detaching the cells with 2 mL PBS/EDTA/trypsin and transferring them into canonical tubes containing 8 mL 10% DMEM. ..

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    Millipore vero cells pedv positive intestinal
    (A) Electron micrograph of a <t>PEDV-infected</t> <t>Vero</t> cell. PEDV particles (arrow heads) on the cell surface and inside a vesicle (arrow) of infected Vero cell. Scale bar = 200 nm. (B) Electron microscopy images of PEDV virions from cell culture media of Vero cells infected with different PEDV strains: PC22A (highly virulent), PC177 (197 aa S deletion) and Iowa106 (sample #PV39, S INDEL). Scale bars = 100 nm.
    Vero Cells Pedv Positive Intestinal, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    (A) Electron micrograph of a PEDV-infected Vero cell. PEDV particles (arrow heads) on the cell surface and inside a vesicle (arrow) of infected Vero cell. Scale bar = 200 nm. (B) Electron microscopy images of PEDV virions from cell culture media of Vero cells infected with different PEDV strains: PC22A (highly virulent), PC177 (197 aa S deletion) and Iowa106 (sample #PV39, S INDEL). Scale bars = 100 nm.

    Journal: Veterinary Microbiology

    Article Title: Cell culture isolation and sequence analysis of genetically diverse US porcine epidemic diarrhea virus strains including a novel strain with a large deletion in the spike gene

    doi: 10.1016/j.vetmic.2014.08.012

    Figure Lengend Snippet: (A) Electron micrograph of a PEDV-infected Vero cell. PEDV particles (arrow heads) on the cell surface and inside a vesicle (arrow) of infected Vero cell. Scale bar = 200 nm. (B) Electron microscopy images of PEDV virions from cell culture media of Vero cells infected with different PEDV strains: PC22A (highly virulent), PC177 (197 aa S deletion) and Iowa106 (sample #PV39, S INDEL). Scale bars = 100 nm.

    Article Snippet: 2.4 Preparation of PEDV inoculum for Vero cells PEDV-positive intestinal or fecal samples were diluted 10-fold in Dulbecco's Phosphate buffered Saline without Mg2+ and Ca2+ [PBS (−), Sigma, St. Louis, MO), and were vortexed briefly followed by centrifugation at 10,000 × g for 3 min at 4 °C.

    Techniques: Infection, Electron Microscopy, Cell Culture

    PEDV strains, PC22A (highly virulent), PC177 (197 aa S deletion) and Iowa106 (sample #PV39, S INDEL) replicated in Vero cells. Upper and lower panels show light and the corresponding immunofluorescent microscope images, respectively, of Vero cells infected with the PEDV strains. PEDV-infected Vero cells were fixed at 2 PID. PEDV and nuclei were detected with mouse anti-PEDV N protein monoclonal antibody #6-29 and 4′,6-diamidino-2-phenylindole (DAPI), respectively. The infected cells showed obvious syncytial cells containing different numbers of nuclei (green for PEDV antigens, and blue for nuclei).

    Journal: Veterinary Microbiology

    Article Title: Cell culture isolation and sequence analysis of genetically diverse US porcine epidemic diarrhea virus strains including a novel strain with a large deletion in the spike gene

    doi: 10.1016/j.vetmic.2014.08.012

    Figure Lengend Snippet: PEDV strains, PC22A (highly virulent), PC177 (197 aa S deletion) and Iowa106 (sample #PV39, S INDEL) replicated in Vero cells. Upper and lower panels show light and the corresponding immunofluorescent microscope images, respectively, of Vero cells infected with the PEDV strains. PEDV-infected Vero cells were fixed at 2 PID. PEDV and nuclei were detected with mouse anti-PEDV N protein monoclonal antibody #6-29 and 4′,6-diamidino-2-phenylindole (DAPI), respectively. The infected cells showed obvious syncytial cells containing different numbers of nuclei (green for PEDV antigens, and blue for nuclei).

    Article Snippet: 2.4 Preparation of PEDV inoculum for Vero cells PEDV-positive intestinal or fecal samples were diluted 10-fold in Dulbecco's Phosphate buffered Saline without Mg2+ and Ca2+ [PBS (−), Sigma, St. Louis, MO), and were vortexed briefly followed by centrifugation at 10,000 × g for 3 min at 4 °C.

    Techniques: Microscopy, Infection

    FACS analyses on A549, fibroblasts, and Vero cells for the expression of CD55, CD59, and CD46. Detection of membrane-anchored complement regulator protein (mCRP) expression on ZIKV-producing cells by FACS analysis. Cells were stained with anti-human CD46, CD55, or CD59 (dark grey histogram) or isotype-matched Ab (light gray histogram).

    Journal: Viruses

    Article Title: Incorporation of CD55 into the Zika Viral Envelope Contributes to Its Stability against Human Complement

    doi: 10.3390/v13030510

    Figure Lengend Snippet: FACS analyses on A549, fibroblasts, and Vero cells for the expression of CD55, CD59, and CD46. Detection of membrane-anchored complement regulator protein (mCRP) expression on ZIKV-producing cells by FACS analysis. Cells were stained with anti-human CD46, CD55, or CD59 (dark grey histogram) or isotype-matched Ab (light gray histogram).

    Article Snippet: Flow Cytometric Analysis For the detection of membrane-bound complement regulatory proteins (mCRPs), virus-producing cells were immune-stained as follow: Vero cells, primary Fibroblasts, and A549 cells (5 × 105 cells/tube) were harvested by detaching with Accutase (Sigma, Vienna Austria), washed twice with buffer containing 2% FBS, 0.2% EDTA in PBS, and incubated with 10 µg/mL anti-CD59, anti-CD55, anti-CD46 or isotope control Abs for 1 h at 4 °C.

    Techniques: FACS, Expressing, Staining

    Serum stability of Zika virus (ZIKV) is affected by cell-line-dependent features. ( A ) ZIKV Insect , ZIKV A549 , ZIKV Fibro, and ZIKV Vero were incubated with increasing amounts of active or heat-inactivated human serum. ( B ) For analysis of the different viral preparations, the input virus was normalized on the amount of virus in heat-inactivated normal human serum (hiNHS). The graph shows the stability of ZIKV derived from different cells in 50%NHS. In both sets of experiments, virus-serum mixtures were incubated for 1 h at 37 °C, then diluted 10-fold, titrated on 12-well plates of overnight-plated Vero cells and incubated for 1 h before plaque agarose was overlaid. Plaques were visualized four days post infection using crystal violet staining. All assays were performed in triplicate, and the error bars show standard deviations. A 95% significance level ( p

    Journal: Viruses

    Article Title: Incorporation of CD55 into the Zika Viral Envelope Contributes to Its Stability against Human Complement

    doi: 10.3390/v13030510

    Figure Lengend Snippet: Serum stability of Zika virus (ZIKV) is affected by cell-line-dependent features. ( A ) ZIKV Insect , ZIKV A549 , ZIKV Fibro, and ZIKV Vero were incubated with increasing amounts of active or heat-inactivated human serum. ( B ) For analysis of the different viral preparations, the input virus was normalized on the amount of virus in heat-inactivated normal human serum (hiNHS). The graph shows the stability of ZIKV derived from different cells in 50%NHS. In both sets of experiments, virus-serum mixtures were incubated for 1 h at 37 °C, then diluted 10-fold, titrated on 12-well plates of overnight-plated Vero cells and incubated for 1 h before plaque agarose was overlaid. Plaques were visualized four days post infection using crystal violet staining. All assays were performed in triplicate, and the error bars show standard deviations. A 95% significance level ( p

    Article Snippet: Flow Cytometric Analysis For the detection of membrane-bound complement regulatory proteins (mCRPs), virus-producing cells were immune-stained as follow: Vero cells, primary Fibroblasts, and A549 cells (5 × 105 cells/tube) were harvested by detaching with Accutase (Sigma, Vienna Austria), washed twice with buffer containing 2% FBS, 0.2% EDTA in PBS, and incubated with 10 µg/mL anti-CD59, anti-CD55, anti-CD46 or isotope control Abs for 1 h at 4 °C.

    Techniques: Incubation, Derivative Assay, Infection, Staining

    Western blot of ZIKV lysates confirms CD55 uptake. ZIKV-containing supernatants (concentrated) of infected A549, Vero, or Insect cells were characterized for CD55 incorporation by Western blotting. Samples were incubated with anti-human CD55 antibody. As load control, anti-ZIKV envelope was used. A representative Western blot analysis is shown.

    Journal: Viruses

    Article Title: Incorporation of CD55 into the Zika Viral Envelope Contributes to Its Stability against Human Complement

    doi: 10.3390/v13030510

    Figure Lengend Snippet: Western blot of ZIKV lysates confirms CD55 uptake. ZIKV-containing supernatants (concentrated) of infected A549, Vero, or Insect cells were characterized for CD55 incorporation by Western blotting. Samples were incubated with anti-human CD55 antibody. As load control, anti-ZIKV envelope was used. A representative Western blot analysis is shown.

    Article Snippet: Flow Cytometric Analysis For the detection of membrane-bound complement regulatory proteins (mCRPs), virus-producing cells were immune-stained as follow: Vero cells, primary Fibroblasts, and A549 cells (5 × 105 cells/tube) were harvested by detaching with Accutase (Sigma, Vienna Austria), washed twice with buffer containing 2% FBS, 0.2% EDTA in PBS, and incubated with 10 µg/mL anti-CD59, anti-CD55, anti-CD46 or isotope control Abs for 1 h at 4 °C.

    Techniques: Western Blot, Infection, Incubation

    Capture ELISA indicates CD55 incorporation in ZIKV envelope. Graphs depict fold changes of RNA genome copies of captured ZIKV, analyzed by RT-PCR. Intact virions ZIKV A549 , ZIKV Insect , ZIKV Fibro , or ZIKV Vero were incubated overnight on pre-coated ELISA plates, bearing antibodies against human CD46, CD55, and CD59. ZIKV envelope served as a positive control. Isotype control was used as a negative control. After extensive washing steps, in order to remove unbound virions, ZIKV particles were lysed and RNA was extracted. The captured copy number of ZIKV genomes was characterized by determining the threshold-cycles by qPCR. Experiments were done two times in duplicates, and the error bars show standard deviations. Data were analyzed by ANOVA followed by Sidak’s multiple comparisons. A 95% significance level ( p

    Journal: Viruses

    Article Title: Incorporation of CD55 into the Zika Viral Envelope Contributes to Its Stability against Human Complement

    doi: 10.3390/v13030510

    Figure Lengend Snippet: Capture ELISA indicates CD55 incorporation in ZIKV envelope. Graphs depict fold changes of RNA genome copies of captured ZIKV, analyzed by RT-PCR. Intact virions ZIKV A549 , ZIKV Insect , ZIKV Fibro , or ZIKV Vero were incubated overnight on pre-coated ELISA plates, bearing antibodies against human CD46, CD55, and CD59. ZIKV envelope served as a positive control. Isotype control was used as a negative control. After extensive washing steps, in order to remove unbound virions, ZIKV particles were lysed and RNA was extracted. The captured copy number of ZIKV genomes was characterized by determining the threshold-cycles by qPCR. Experiments were done two times in duplicates, and the error bars show standard deviations. Data were analyzed by ANOVA followed by Sidak’s multiple comparisons. A 95% significance level ( p

    Article Snippet: Flow Cytometric Analysis For the detection of membrane-bound complement regulatory proteins (mCRPs), virus-producing cells were immune-stained as follow: Vero cells, primary Fibroblasts, and A549 cells (5 × 105 cells/tube) were harvested by detaching with Accutase (Sigma, Vienna Austria), washed twice with buffer containing 2% FBS, 0.2% EDTA in PBS, and incubated with 10 µg/mL anti-CD59, anti-CD55, anti-CD46 or isotope control Abs for 1 h at 4 °C.

    Techniques: Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Incubation, Positive Control, Negative Control, Real-time Polymerase Chain Reaction

    Effect of IgM on complement activation. To investigate whether natural serum IgMs affect the complement activation of human cell-derived ZIKV, anti-human IgM blocking antibodies were incubated in 50% NHS or hiNHS for 30 min on ice, before ZIKV A549 or ZIKV Fibro were added. The experimental conditions are indicated by “x”. As positive control, ZIKV insect was included [ 13 ]. After incubation for 1 h at 37 °C, the virus–serum mixture was serial diluted and titrated on 12-well plates of overnight-plated Vero cells. One hour after incubation at 37 °C, plaque agarose was overlaid. Viral concentration was determined and calculated four days post-infection using crystal violet staining. All virus lysis experiments were conducted in triplicate, and the error bars show standard deviations. A 95% significance level ( p

    Journal: Viruses

    Article Title: Incorporation of CD55 into the Zika Viral Envelope Contributes to Its Stability against Human Complement

    doi: 10.3390/v13030510

    Figure Lengend Snippet: Effect of IgM on complement activation. To investigate whether natural serum IgMs affect the complement activation of human cell-derived ZIKV, anti-human IgM blocking antibodies were incubated in 50% NHS or hiNHS for 30 min on ice, before ZIKV A549 or ZIKV Fibro were added. The experimental conditions are indicated by “x”. As positive control, ZIKV insect was included [ 13 ]. After incubation for 1 h at 37 °C, the virus–serum mixture was serial diluted and titrated on 12-well plates of overnight-plated Vero cells. One hour after incubation at 37 °C, plaque agarose was overlaid. Viral concentration was determined and calculated four days post-infection using crystal violet staining. All virus lysis experiments were conducted in triplicate, and the error bars show standard deviations. A 95% significance level ( p

    Article Snippet: Flow Cytometric Analysis For the detection of membrane-bound complement regulatory proteins (mCRPs), virus-producing cells were immune-stained as follow: Vero cells, primary Fibroblasts, and A549 cells (5 × 105 cells/tube) were harvested by detaching with Accutase (Sigma, Vienna Austria), washed twice with buffer containing 2% FBS, 0.2% EDTA in PBS, and incubated with 10 µg/mL anti-CD59, anti-CD55, anti-CD46 or isotope control Abs for 1 h at 4 °C.

    Techniques: Activation Assay, Derivative Assay, Blocking Assay, Incubation, Positive Control, Concentration Assay, Infection, Staining, Lysis

    Effect of cholesterol depletion from cells on the course of HSV entry. Vero cells were infected with HSV-1 (KOS/tk12). At various times following infection (indicated on the abscissa), cells were treated for 30 min with MβCD. The drug was washed out, and medium or cholesterol (chol) was added. After 30 min, the cholesterol was removed, and infection was allowed to proceed. β-Galactosidase activity at 6 h postinfection was determined. The results shown are representative of three independent experiments, and 100% entry represents the β-galactosidase activity at 6 h in the absence of MβCD.

    Journal: Journal of Virology

    Article Title: Specific Association of Glycoprotein B with Lipid Rafts during Herpes Simplex Virus Entry

    doi: 10.1128/JVI.77.17.9542-9552.2003

    Figure Lengend Snippet: Effect of cholesterol depletion from cells on the course of HSV entry. Vero cells were infected with HSV-1 (KOS/tk12). At various times following infection (indicated on the abscissa), cells were treated for 30 min with MβCD. The drug was washed out, and medium or cholesterol (chol) was added. After 30 min, the cholesterol was removed, and infection was allowed to proceed. β-Galactosidase activity at 6 h postinfection was determined. The results shown are representative of three independent experiments, and 100% entry represents the β-galactosidase activity at 6 h in the absence of MβCD.

    Article Snippet: In replenishment experiments, Vero cells were first treated with 7.5 mM MβCD for 30 min. MβCD was washed out as before, and various amounts of water-soluble cholesterol balanced with MβCD (Sigma) in DMEM without serum were added.

    Techniques: Infection, Activity Assay

    Effect of cholesterol depletion or chelation on HSV entry. Vero cells (A and B) and B78 A10 and C10 cells (C) were treated with increasing concentrations of MβCD (A and C) or nystatin (B). The drugs were washed out, and the cells were infected with HSV-1 (KOS/tk12) and assayed for β-galactosidase activity at 6 h postinfection. Results presented in A and B are the mean of three independent experiments done in duplicate, with standard deviations. The results shown in panel C are representative of three experiments. A value of 100% entry represents the β-galactosidase activity at 6 h in the absence of MβCD.

    Journal: Journal of Virology

    Article Title: Specific Association of Glycoprotein B with Lipid Rafts during Herpes Simplex Virus Entry

    doi: 10.1128/JVI.77.17.9542-9552.2003

    Figure Lengend Snippet: Effect of cholesterol depletion or chelation on HSV entry. Vero cells (A and B) and B78 A10 and C10 cells (C) were treated with increasing concentrations of MβCD (A and C) or nystatin (B). The drugs were washed out, and the cells were infected with HSV-1 (KOS/tk12) and assayed for β-galactosidase activity at 6 h postinfection. Results presented in A and B are the mean of three independent experiments done in duplicate, with standard deviations. The results shown in panel C are representative of three experiments. A value of 100% entry represents the β-galactosidase activity at 6 h in the absence of MβCD.

    Article Snippet: In replenishment experiments, Vero cells were first treated with 7.5 mM MβCD for 30 min. MβCD was washed out as before, and various amounts of water-soluble cholesterol balanced with MβCD (Sigma) in DMEM without serum were added.

    Techniques: Infection, Activity Assay

    Effect of cholesterol depletion from cells on expression of nectin-1 and on attachment and replication of HSV. (A) C10 cells were untreated (−) or treated (+) with 30 mM MβCD for 30 min. The drug was washed out, and HSV-1 (KOS) was allowed to bind to the cells for 1 h at 4°C. Total cell proteins were extracted, resolved by SDS-PAGE, and probed with anti-nectin-1 MAb CK6 (upper panel) or anti-gD MAb DL6 (lower panel). (B) C10 cells were treated with increasing concentrations of MβCD (indicated at the right), then stained with anti-nectin-1 MAb CK41 directly labeled with phycoerythrin, and analyzed by FACS. The control (open area) represents the fluorescence of unstained and drug untreated cells. (C) Vero cells were treated with increasing concentrations of MβCD. The drug was washed out, and the cells were infected with HSV1 (KOS) or vesicular stomatitis virus (VSV) for 24 h at 37°C. Then the cells were fixed, and plaques werevisualized by immunoperoxidase staining (HSV) or crystal violet staining (vesicular stomatitis virus).

    Journal: Journal of Virology

    Article Title: Specific Association of Glycoprotein B with Lipid Rafts during Herpes Simplex Virus Entry

    doi: 10.1128/JVI.77.17.9542-9552.2003

    Figure Lengend Snippet: Effect of cholesterol depletion from cells on expression of nectin-1 and on attachment and replication of HSV. (A) C10 cells were untreated (−) or treated (+) with 30 mM MβCD for 30 min. The drug was washed out, and HSV-1 (KOS) was allowed to bind to the cells for 1 h at 4°C. Total cell proteins were extracted, resolved by SDS-PAGE, and probed with anti-nectin-1 MAb CK6 (upper panel) or anti-gD MAb DL6 (lower panel). (B) C10 cells were treated with increasing concentrations of MβCD (indicated at the right), then stained with anti-nectin-1 MAb CK41 directly labeled with phycoerythrin, and analyzed by FACS. The control (open area) represents the fluorescence of unstained and drug untreated cells. (C) Vero cells were treated with increasing concentrations of MβCD. The drug was washed out, and the cells were infected with HSV1 (KOS) or vesicular stomatitis virus (VSV) for 24 h at 37°C. Then the cells were fixed, and plaques werevisualized by immunoperoxidase staining (HSV) or crystal violet staining (vesicular stomatitis virus).

    Article Snippet: In replenishment experiments, Vero cells were first treated with 7.5 mM MβCD for 30 min. MβCD was washed out as before, and various amounts of water-soluble cholesterol balanced with MβCD (Sigma) in DMEM without serum were added.

    Techniques: Expressing, SDS Page, Staining, Labeling, FACS, Fluorescence, Infection, Immunoperoxidase Staining

    Effect of cholesterol replenishment on HSV entry. Vero cells were untreated (left, rectangles) or treated (right, rectangles) with 7.5 mM MβCD for 30 min. MβCD was washed out, and various amounts of cholesterol in MβCD were added. After 30 min, the cholesterol was removed, and the cells were infected with HSV-1 (KOS/tk12) and assayed for β-galactosidase activity at 6 h postinfection. The results presented are representative of two independent experiments done in triplicate.

    Journal: Journal of Virology

    Article Title: Specific Association of Glycoprotein B with Lipid Rafts during Herpes Simplex Virus Entry

    doi: 10.1128/JVI.77.17.9542-9552.2003

    Figure Lengend Snippet: Effect of cholesterol replenishment on HSV entry. Vero cells were untreated (left, rectangles) or treated (right, rectangles) with 7.5 mM MβCD for 30 min. MβCD was washed out, and various amounts of cholesterol in MβCD were added. After 30 min, the cholesterol was removed, and the cells were infected with HSV-1 (KOS/tk12) and assayed for β-galactosidase activity at 6 h postinfection. The results presented are representative of two independent experiments done in triplicate.

    Article Snippet: In replenishment experiments, Vero cells were first treated with 7.5 mM MβCD for 30 min. MβCD was washed out as before, and various amounts of water-soluble cholesterol balanced with MβCD (Sigma) in DMEM without serum were added.

    Techniques: Infection, Activity Assay

    Growth of NS 116 -GFP/AT and NS 116 -GFP/AE Viruses in Different Cell Lines Vero, B16f1, PANC-1, A375, or CaCo2 1 × 10 5 cells were infected with either NS 116 -GFP/AT or NS 116 -GFP/AE viruses at an MOI of 0.01 and cultivated in the presence trypsin or neutrophil elastase, respectively. Supernatant was collected 12, 24, 48, and 72 hr post-infection. Data represent means + SEM (n = 3).

    Journal: Molecular Therapy Oncolytics

    Article Title: Targeting an Oncolytic Influenza A Virus to Tumor Tissue by Elastase

    doi: 10.1016/j.omto.2017.09.002

    Figure Lengend Snippet: Growth of NS 116 -GFP/AT and NS 116 -GFP/AE Viruses in Different Cell Lines Vero, B16f1, PANC-1, A375, or CaCo2 1 × 10 5 cells were infected with either NS 116 -GFP/AT or NS 116 -GFP/AE viruses at an MOI of 0.01 and cultivated in the presence trypsin or neutrophil elastase, respectively. Supernatant was collected 12, 24, 48, and 72 hr post-infection. Data represent means + SEM (n = 3).

    Article Snippet: Trypsin-dependent NS116 -GFP/AT or elastase-dependent NS1116 -GFP/AE viruses were propagated at 37°C in Vero cells infected with MOI of 0.01 in the presence of either 1 μg L-1-Tosylamide-2-phenylethyl chloromethyl ketone (TPCK)-treated trypsin (Sigma Aldrich) ml−1 or 10 μg human neutrophil elastase (Serva Electrophoresis, Heidelberg, Germany) ml−1 .

    Techniques: Infection

    Growth Kinetic of Modified NS 116 -GFP/AE Virus A 24-hr-old monolayer of Vero cells was infected with NS 116 -GFP/AE virus in the presence of either pancreatic elastase (p-elastase) or neutrophil elastase (n-elastase). A viral titer was determined by TCID 50 in Vero cells at the indicated time points + SEM (n = 6).

    Journal: Molecular Therapy Oncolytics

    Article Title: Targeting an Oncolytic Influenza A Virus to Tumor Tissue by Elastase

    doi: 10.1016/j.omto.2017.09.002

    Figure Lengend Snippet: Growth Kinetic of Modified NS 116 -GFP/AE Virus A 24-hr-old monolayer of Vero cells was infected with NS 116 -GFP/AE virus in the presence of either pancreatic elastase (p-elastase) or neutrophil elastase (n-elastase). A viral titer was determined by TCID 50 in Vero cells at the indicated time points + SEM (n = 6).

    Article Snippet: Trypsin-dependent NS116 -GFP/AT or elastase-dependent NS1116 -GFP/AE viruses were propagated at 37°C in Vero cells infected with MOI of 0.01 in the presence of either 1 μg L-1-Tosylamide-2-phenylethyl chloromethyl ketone (TPCK)-treated trypsin (Sigma Aldrich) ml−1 or 10 μg human neutrophil elastase (Serva Electrophoresis, Heidelberg, Germany) ml−1 .

    Techniques: Modification, Infection

    Growth Properties of NS 116 -GFP/AE and NS 116 -GFP/AT Virus in Vero Cells in Co-culture System with Human Neutrophils or Supernatant Supplemented with Neutrophil Elastase or Trypsin (A) Vero cells were infected at an MOI of 0.01 under indicated condition and viral titer was determined by TCID 50 24 hr post-infection. Data represent means + SEM (n = 3). (B) Virus induced GFP expression, which indicates viral growth. The pictures of infected Vero cells were taken 48 hr post-infection using an AxioCam ICc3 Rev.2-3 camera, with a magnification of 200×. Experiments were carried out three times with neutrophils of another proband, and the corresponding results are provided. Scale bar, 100 μm.

    Journal: Molecular Therapy Oncolytics

    Article Title: Targeting an Oncolytic Influenza A Virus to Tumor Tissue by Elastase

    doi: 10.1016/j.omto.2017.09.002

    Figure Lengend Snippet: Growth Properties of NS 116 -GFP/AE and NS 116 -GFP/AT Virus in Vero Cells in Co-culture System with Human Neutrophils or Supernatant Supplemented with Neutrophil Elastase or Trypsin (A) Vero cells were infected at an MOI of 0.01 under indicated condition and viral titer was determined by TCID 50 24 hr post-infection. Data represent means + SEM (n = 3). (B) Virus induced GFP expression, which indicates viral growth. The pictures of infected Vero cells were taken 48 hr post-infection using an AxioCam ICc3 Rev.2-3 camera, with a magnification of 200×. Experiments were carried out three times with neutrophils of another proband, and the corresponding results are provided. Scale bar, 100 μm.

    Article Snippet: Trypsin-dependent NS116 -GFP/AT or elastase-dependent NS1116 -GFP/AE viruses were propagated at 37°C in Vero cells infected with MOI of 0.01 in the presence of either 1 μg L-1-Tosylamide-2-phenylethyl chloromethyl ketone (TPCK)-treated trypsin (Sigma Aldrich) ml−1 or 10 μg human neutrophil elastase (Serva Electrophoresis, Heidelberg, Germany) ml−1 .

    Techniques: Co-Culture Assay, Infection, Expressing