ma 104 cells  (ATCC)


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    Name:
    MA 104 Clone 1
    Description:

    Catalog Number:
    crl-2378.1
    Price:
    None
    Applications:
    This cell line is a suitable transfection host.
    Cell Type:
    epithelial cell
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    Structured Review

    ATCC ma 104 cells
    <t>MA-104</t> cells infected or mock infected with the indicated virus. The presence of the virus was determined by using a monoclonal antibody that detects ASFV protein p30, and it was visualized using an immunoproxidase assay.

    https://www.bioz.com/result/ma 104 cells/product/ATCC
    Average 99 stars, based on 1 article reviews
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    ma 104 cells - by Bioz Stars, 2020-09
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    Images

    1) Product Images from "Identification of a Continuously Stable and Commercially Available Cell Line for the Identification of Infectious African Swine Fever Virus in Clinical Samples"

    Article Title: Identification of a Continuously Stable and Commercially Available Cell Line for the Identification of Infectious African Swine Fever Virus in Clinical Samples

    Journal: Viruses

    doi: 10.3390/v12080820

    MA-104 cells infected or mock infected with the indicated virus. The presence of the virus was determined by using a monoclonal antibody that detects ASFV protein p30, and it was visualized using an immunoproxidase assay.
    Figure Legend Snippet: MA-104 cells infected or mock infected with the indicated virus. The presence of the virus was determined by using a monoclonal antibody that detects ASFV protein p30, and it was visualized using an immunoproxidase assay.

    Techniques Used: Infection

    Comparison of the titration of African Swine Fever Virus (ASFV) isolates in swine macrophages (blue) or MA-104 (orange) cells with real-time PCR. Titrations are expressed in log 10 HA 50 /mL, the error bars are the standard deviation between replicates, and rt-PCR is expressed as L]log 10 /mL, indicating the last dilution that had a positive CT value for ASFV.
    Figure Legend Snippet: Comparison of the titration of African Swine Fever Virus (ASFV) isolates in swine macrophages (blue) or MA-104 (orange) cells with real-time PCR. Titrations are expressed in log 10 HA 50 /mL, the error bars are the standard deviation between replicates, and rt-PCR is expressed as L]log 10 /mL, indicating the last dilution that had a positive CT value for ASFV.

    Techniques Used: Titration, Real-time Polymerase Chain Reaction, Standard Deviation, Reverse Transcription Polymerase Chain Reaction

    MA-104 cells were infected with the indicated virus in the presence of red blood cells. Hemadsorption was observed 24 h after infection.
    Figure Legend Snippet: MA-104 cells were infected with the indicated virus in the presence of red blood cells. Hemadsorption was observed 24 h after infection.

    Techniques Used: Infection

    2) Product Images from "Identification of Nonessential Regions of the nsp2 Replicase Protein of Porcine Reproductive and Respiratory Syndrome Virus Strain VR-2332 for Replication in Cell Culture ▿"

    Article Title: Identification of Nonessential Regions of the nsp2 Replicase Protein of Porcine Reproductive and Respiratory Syndrome Virus Strain VR-2332 for Replication in Cell Culture ▿

    Journal:

    doi: 10.1128/JVI.00562-07

    Utilization of nsp2 to express foreign gene. Viable GFP recombinant virus was generated in MA-104 cells. (A and B) Light (A) and fluorescence (B) (×10) microscopy under the same field showed the fluorescence was associated with the virus-infected
    Figure Legend Snippet: Utilization of nsp2 to express foreign gene. Viable GFP recombinant virus was generated in MA-104 cells. (A and B) Light (A) and fluorescence (B) (×10) microscopy under the same field showed the fluorescence was associated with the virus-infected

    Techniques Used: Recombinant, Generated, Fluorescence, Microscopy, Infection

    3) Product Images from "Identification of a Continuously Stable and Commercially Available Cell Line for the Identification of Infectious African Swine Fever Virus in Clinical Samples"

    Article Title: Identification of a Continuously Stable and Commercially Available Cell Line for the Identification of Infectious African Swine Fever Virus in Clinical Samples

    Journal: Viruses

    doi: 10.3390/v12080820

    MA-104 cells infected or mock infected with the indicated virus. The presence of the virus was determined by using a monoclonal antibody that detects ASFV protein p30, and it was visualized using an immunoproxidase assay.
    Figure Legend Snippet: MA-104 cells infected or mock infected with the indicated virus. The presence of the virus was determined by using a monoclonal antibody that detects ASFV protein p30, and it was visualized using an immunoproxidase assay.

    Techniques Used: Infection

    Comparison of the titration of African Swine Fever Virus (ASFV) isolates in swine macrophages (blue) or MA-104 (orange) cells with real-time PCR. Titrations are expressed in log 10 HA 50 /mL, the error bars are the standard deviation between replicates, and rt-PCR is expressed as L]log 10 /mL, indicating the last dilution that had a positive CT value for ASFV.
    Figure Legend Snippet: Comparison of the titration of African Swine Fever Virus (ASFV) isolates in swine macrophages (blue) or MA-104 (orange) cells with real-time PCR. Titrations are expressed in log 10 HA 50 /mL, the error bars are the standard deviation between replicates, and rt-PCR is expressed as L]log 10 /mL, indicating the last dilution that had a positive CT value for ASFV.

    Techniques Used: Titration, Real-time Polymerase Chain Reaction, Standard Deviation, Reverse Transcription Polymerase Chain Reaction

    MA-104 cells were infected with the indicated virus in the presence of red blood cells. Hemadsorption was observed 24 h after infection.
    Figure Legend Snippet: MA-104 cells were infected with the indicated virus in the presence of red blood cells. Hemadsorption was observed 24 h after infection.

    Techniques Used: Infection

    Related Articles

    Modification:

    Article Title: In Vitro Neutralisation of Rotavirus Infection by Two Broadly Specific Recombinant Monovalent Llama-Derived Antibody Fragments
    Article Snippet: .. At the Laboratory of Paediatrics, Erasmus MC, CaCo-2 (ATCC, HTB-37) or MA104 (ATCC, CRL-2378.1) cells were maintained in Dulbecco's Modified Eagle's Medium (DMEM, GibcoBRL, Paisley, Scotland) containing 10% (v/v) foetal calf serum (FBS, Integro, Dieren, The Netherlands), 100 U/ml Penicillin, 100 µg/ml Streptomycin and 1% (v/v) non-essential amino acids (BioWhittacker, Verviers, Belgium) at 37°C and in an atmosphere of 5% CO2 -air. .. To test the neutralising activity of the llama antibody fragment ARP1 and a control antibody (VHH R2), 1.5×104 CaCo-2 cells were plated on heavy Teflon coated microscope slides (∅ 7 mm, Cell-line/Erie Scientific, Portsmouth, NH) as described previously .

    Article Title: Recombinant Rotaviruses Rescued by Reverse Genetics Reveal the Role of NSP5 Hyperphosphorylation in the Assembly of Viral Factories
    Article Snippet: .. MA104 (embryonic African green monkey kidney cells, ATCC CRL-2378.1, from Chlorocebus aethiops ), U2OS (human bone osteosarcoma epithelial cells), Caco-2 (colorectal adenocarcinoma human intestinal epithelial cell line, ATCC HTB-37), and HEK293T (human embryonic kidney epithelial cells, ATCC CRL-3216) cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Life Technologies) supplemented with 10% fetal bovine serum (FBS) (Life Technologies) and 50 μg/ml gentamicin (Biochrom AG). .. MA104-NSP5-EGFP (MA-NSP5-EGFP) cells ( ) were cultured in DMEM supplemented with 10% FBS (Life Technologies), 50 μg/ml gentamicin (Biochrom AG), and 1 mg/ml Geneticin (Gibco-BRL, Life Technologies).

    other:

    Article Title: Identification of a Continuously Stable and Commercially Available Cell Line for the Identification of Infectious African Swine Fever Virus in Clinical Samples
    Article Snippet: Historically, MA-104 cells contained a mix culture from two different species of monkeys.

    Cell Culture:

    Article Title: Rotavirus VP3 targets MAVS for degradation to inhibit type III interferon expression in intestinal epithelial cells
    Article Snippet: .. Cells African Green Monkey kidney epithelial cell lines MA104 (CRL-2378.1) and COS-7 (CRL-1651) were originally obtained from American Type Culture Collection (ATCC) and cultured in complete M199 medium. .. Human embryonic kidney cell line HEK293 (CRL-1573) and mouse embryonic fibroblast cell line NIH3T3 (CRL-1658) were obtained from ATCC and cultured in complete DMEM medium.

    Article Title: Recombinant Rotaviruses Rescued by Reverse Genetics Reveal the Role of NSP5 Hyperphosphorylation in the Assembly of Viral Factories
    Article Snippet: .. MA104 (embryonic African green monkey kidney cells, ATCC CRL-2378.1, from Chlorocebus aethiops ), U2OS (human bone osteosarcoma epithelial cells), Caco-2 (colorectal adenocarcinoma human intestinal epithelial cell line, ATCC HTB-37), and HEK293T (human embryonic kidney epithelial cells, ATCC CRL-3216) cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Life Technologies) supplemented with 10% fetal bovine serum (FBS) (Life Technologies) and 50 μg/ml gentamicin (Biochrom AG). .. MA104-NSP5-EGFP (MA-NSP5-EGFP) cells ( ) were cultured in DMEM supplemented with 10% FBS (Life Technologies), 50 μg/ml gentamicin (Biochrom AG), and 1 mg/ml Geneticin (Gibco-BRL, Life Technologies).

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    ATCC confluent ma104 cell monolayers
    SB105-A10 does not inhibit the infectivity of a human rotavirus. <t>MA104</t> cells grown in 96-well plates were infected for 1 h at 37°C with a human rotavirus (strain Wa) in the presence of increasing concentrations of SB105-A10. Results shown are
    Confluent Ma104 Cell Monolayers, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/confluent ma104 cell monolayers/product/ATCC
    Average 99 stars, based on 14 article reviews
    Price from $9.99 to $1999.99
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    SB105-A10 does not inhibit the infectivity of a human rotavirus. MA104 cells grown in 96-well plates were infected for 1 h at 37°C with a human rotavirus (strain Wa) in the presence of increasing concentrations of SB105-A10. Results shown are

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Identification of a Dendrimeric Heparan Sulfate-Binding Peptide That Inhibits Infectivity of Genital Types of Human Papillomaviruses ▿

    doi: 10.1128/AAC.00471-10

    Figure Lengend Snippet: SB105-A10 does not inhibit the infectivity of a human rotavirus. MA104 cells grown in 96-well plates were infected for 1 h at 37°C with a human rotavirus (strain Wa) in the presence of increasing concentrations of SB105-A10. Results shown are

    Article Snippet: Confluent MA104 cell monolayers in a 96-well plate were washed twice with MEM and then infected with 1,200 PFU of human rotavirus strain Wa (ATCC VR-2018) for 1 h at 37°C in the presence or absence of SB105-A10.

    Techniques: Infection

    Electron microscopy of RV-infected cells treated with ML. High-definition electron microscopy of noninfected (NI) and RV-infected (OSU; MOI, 100 VFU/ml) MA104 cells untreated (DMSO) or treated with ML (20 μM) from 1 hpi. At 6 hpi, the cells were fixed with glutaraldehyde and processed for transmission electron microscopy. V, viroplasms; Nu, nucleus, ER, endoplasmic reticulum; Gg, Golgi complex; Vc, vacuoles; P h , phagosomes; CM, cell membrane; the thin arrows indicate the endoplasmic reticulum membrane surrounding viroplasms; the large arrowheads indicate viral particles.

    Journal: Journal of Virology

    Article Title: Identification of a Small Molecule That Compromises the Structural Integrity of Viroplasms and Rotavirus Double-Layered Particles

    doi: 10.1128/JVI.01943-17

    Figure Lengend Snippet: Electron microscopy of RV-infected cells treated with ML. High-definition electron microscopy of noninfected (NI) and RV-infected (OSU; MOI, 100 VFU/ml) MA104 cells untreated (DMSO) or treated with ML (20 μM) from 1 hpi. At 6 hpi, the cells were fixed with glutaraldehyde and processed for transmission electron microscopy. V, viroplasms; Nu, nucleus, ER, endoplasmic reticulum; Gg, Golgi complex; Vc, vacuoles; P h , phagosomes; CM, cell membrane; the thin arrows indicate the endoplasmic reticulum membrane surrounding viroplasms; the large arrowheads indicate viral particles.

    Article Snippet: MA104 cells (embryonic African green monkey kidney cells; ATCC CRL-2378) were grown in Dulbecco's modified Eagle's medium (DMEM) (Life Technologies) containing 10% fetal bovine serum (Life Technologies) and 50 μg/ml gentamicin (Biochrom AG).

    Techniques: Electron Microscopy, Infection, Transmission Assay

    NSP5 dephosphorylation caused by ML-mediated viroplasm disruption. Shown are Western blot and confocal immunofluorescence analyses with the indicated antibodies of OSU-infected (25 VFU/cell) MA104 cells treated with 10 μM ML and/or 0.5 μM okadaic acid (OA) or DMSO for the indicated times. Scale bars, 5 μm.

    Journal: Journal of Virology

    Article Title: Identification of a Small Molecule That Compromises the Structural Integrity of Viroplasms and Rotavirus Double-Layered Particles

    doi: 10.1128/JVI.01943-17

    Figure Lengend Snippet: NSP5 dephosphorylation caused by ML-mediated viroplasm disruption. Shown are Western blot and confocal immunofluorescence analyses with the indicated antibodies of OSU-infected (25 VFU/cell) MA104 cells treated with 10 μM ML and/or 0.5 μM okadaic acid (OA) or DMSO for the indicated times. Scale bars, 5 μm.

    Article Snippet: MA104 cells (embryonic African green monkey kidney cells; ATCC CRL-2378) were grown in Dulbecco's modified Eagle's medium (DMEM) (Life Technologies) containing 10% fetal bovine serum (Life Technologies) and 50 μg/ml gentamicin (Biochrom AG).

    Techniques: De-Phosphorylation Assay, Western Blot, Immunofluorescence, Infection

    VP6 in RV-infected cells. Shown is confocal immunofluorescence of MA104 cells infected with either OSU or SA11 (MOI, 25 VFU/cell) and transfected with siRNAs specific for SA11 VP6 or OSU VP6 or with a nontargeting siRNA (siNT) (A) or treated with 10 μM ML or DMSO (B). At the indicated times postinfection, viroplasms were visualized with anti-NSP5 antibody (red) and VP6 with MAb 4B2D2 (green). Scale bars, 5 μm.

    Journal: Journal of Virology

    Article Title: Identification of a Small Molecule That Compromises the Structural Integrity of Viroplasms and Rotavirus Double-Layered Particles

    doi: 10.1128/JVI.01943-17

    Figure Lengend Snippet: VP6 in RV-infected cells. Shown is confocal immunofluorescence of MA104 cells infected with either OSU or SA11 (MOI, 25 VFU/cell) and transfected with siRNAs specific for SA11 VP6 or OSU VP6 or with a nontargeting siRNA (siNT) (A) or treated with 10 μM ML or DMSO (B). At the indicated times postinfection, viroplasms were visualized with anti-NSP5 antibody (red) and VP6 with MAb 4B2D2 (green). Scale bars, 5 μm.

    Article Snippet: MA104 cells (embryonic African green monkey kidney cells; ATCC CRL-2378) were grown in Dulbecco's modified Eagle's medium (DMEM) (Life Technologies) containing 10% fetal bovine serum (Life Technologies) and 50 μg/ml gentamicin (Biochrom AG).

    Techniques: Infection, Immunofluorescence, Transfection

    Effect of ML on RV VP6. (A) VP6-VP2 interaction. Shown is Western blot analysis with anti-VP2 and anti-VP6 antibodies of immunoprecipitates (IP) obtained with anti-VP6 MAb RV138 from extracts of MA104 cells transfected with VP6 and VP2 and treated for 5 h with 10 μM ML or DMSO. The inhibitor (200 μM) was maintained during cell lysis and incubation with the precipitating antibody. The numbers on the left are kilodaltons. (B) VP6 trimer stability. (Left) Western blot analysis with anti-VP6 antibody of nonboiled extracts from MA104 cells infected with a recombinant vaccinia virus expressing VP6 (VVVP6) and treated with 10 μM ML or DMSO from 1 to 7 hpi. (Right) Western blot analysis of nonboiled extracts from cells infected with OSU (MOI, 25 VFU/cell) and treated with 10 μM ML or DMSO from 1 to 5 hpi. The numbers on the right are kilodaltons. (C) Confocal immunofluorescence (IF) analysis with the anti-VP6 MAb 4B2D2 of MA104 cells overexpressing VP6 (infected with VVVP6) and treated with 10 μM ML or DMSO from 1 to 7 hpi. The arrow indicates a VP6 higher-order structure observed in the absence of other RV proteins. Scale bars, 5 μm. (D) Representative images of VP6 tubes and spheres visualized by negative-staining electron microscopy after treatment with 25 μM ML for 4 h at 37°C. (E) Interaction of VP6 with ML evaluated by nanoscale thermophoresis. The fraction of Cys- or Lys-labeled VP6 bound to ML was plotted against increasing concentrations of the inhibitor. The data were fitted with two state equations, and an EC 50 of 294 ± 62 μM was calculated as the average of the results of three independent measurements. The error bars indicate standard deviations.

    Journal: Journal of Virology

    Article Title: Identification of a Small Molecule That Compromises the Structural Integrity of Viroplasms and Rotavirus Double-Layered Particles

    doi: 10.1128/JVI.01943-17

    Figure Lengend Snippet: Effect of ML on RV VP6. (A) VP6-VP2 interaction. Shown is Western blot analysis with anti-VP2 and anti-VP6 antibodies of immunoprecipitates (IP) obtained with anti-VP6 MAb RV138 from extracts of MA104 cells transfected with VP6 and VP2 and treated for 5 h with 10 μM ML or DMSO. The inhibitor (200 μM) was maintained during cell lysis and incubation with the precipitating antibody. The numbers on the left are kilodaltons. (B) VP6 trimer stability. (Left) Western blot analysis with anti-VP6 antibody of nonboiled extracts from MA104 cells infected with a recombinant vaccinia virus expressing VP6 (VVVP6) and treated with 10 μM ML or DMSO from 1 to 7 hpi. (Right) Western blot analysis of nonboiled extracts from cells infected with OSU (MOI, 25 VFU/cell) and treated with 10 μM ML or DMSO from 1 to 5 hpi. The numbers on the right are kilodaltons. (C) Confocal immunofluorescence (IF) analysis with the anti-VP6 MAb 4B2D2 of MA104 cells overexpressing VP6 (infected with VVVP6) and treated with 10 μM ML or DMSO from 1 to 7 hpi. The arrow indicates a VP6 higher-order structure observed in the absence of other RV proteins. Scale bars, 5 μm. (D) Representative images of VP6 tubes and spheres visualized by negative-staining electron microscopy after treatment with 25 μM ML for 4 h at 37°C. (E) Interaction of VP6 with ML evaluated by nanoscale thermophoresis. The fraction of Cys- or Lys-labeled VP6 bound to ML was plotted against increasing concentrations of the inhibitor. The data were fitted with two state equations, and an EC 50 of 294 ± 62 μM was calculated as the average of the results of three independent measurements. The error bars indicate standard deviations.

    Article Snippet: MA104 cells (embryonic African green monkey kidney cells; ATCC CRL-2378) were grown in Dulbecco's modified Eagle's medium (DMEM) (Life Technologies) containing 10% fetal bovine serum (Life Technologies) and 50 μg/ml gentamicin (Biochrom AG).

    Techniques: Western Blot, Transfection, Lysis, Incubation, Infection, Recombinant, Expressing, Immunofluorescence, Negative Staining, Electron Microscopy, Labeling

    ML antiviral activity is independent of cellular transcription and protein synthesis. (A) Western blot and confocal immunofluorescence analyses with the indicated antibodies of OSU-infected MA104 cells (25 VFU/cell) fed with EU and treated with 10 μM ML and/or 10 μg/ml actinomycin D (Act D) or DMSO for the indicated times. EU-labeled, newly synthesized RNAs were visualized by reaction with an Alexa-488-conjugated azide (green). Scale bars, 5 μm. (B) Western blot and confocal immunofluorescence analyses with the indicated antibodies of OSU-infected (MOI, 25 VFU/cell) MA104 cells treated with 10 μM ML and/or 10 μg/ml CHX or DMSO for the indicated times. Scale bars, 10 μm.

    Journal: Journal of Virology

    Article Title: Identification of a Small Molecule That Compromises the Structural Integrity of Viroplasms and Rotavirus Double-Layered Particles

    doi: 10.1128/JVI.01943-17

    Figure Lengend Snippet: ML antiviral activity is independent of cellular transcription and protein synthesis. (A) Western blot and confocal immunofluorescence analyses with the indicated antibodies of OSU-infected MA104 cells (25 VFU/cell) fed with EU and treated with 10 μM ML and/or 10 μg/ml actinomycin D (Act D) or DMSO for the indicated times. EU-labeled, newly synthesized RNAs were visualized by reaction with an Alexa-488-conjugated azide (green). Scale bars, 5 μm. (B) Western blot and confocal immunofluorescence analyses with the indicated antibodies of OSU-infected (MOI, 25 VFU/cell) MA104 cells treated with 10 μM ML and/or 10 μg/ml CHX or DMSO for the indicated times. Scale bars, 10 μm.

    Article Snippet: MA104 cells (embryonic African green monkey kidney cells; ATCC CRL-2378) were grown in Dulbecco's modified Eagle's medium (DMEM) (Life Technologies) containing 10% fetal bovine serum (Life Technologies) and 50 μg/ml gentamicin (Biochrom AG).

    Techniques: Activity Assay, Western Blot, Immunofluorescence, Infection, Activated Clotting Time Assay, Labeling, Synthesized

    ML effect on RV replication. (A to E) Western blot and confocal immunofluorescence analyses with the indicated antibodies of RV-infected cells (MOI, 25 VFU/cell) treated with ML at 10 μM, unless otherwise indicated, or with DMSO (D) for the indicated times. Scale bars, 10 μm. TUB, tubulin. (F) Time course of viral progeny yield of OSU-infected (MOI, 25 VFU/cell) MA104 cells treated with 10 μM ML, added at 2 hpi. The data are presented as averages ± standard deviations of the results of three independent experiments. ***, P

    Journal: Journal of Virology

    Article Title: Identification of a Small Molecule That Compromises the Structural Integrity of Viroplasms and Rotavirus Double-Layered Particles

    doi: 10.1128/JVI.01943-17

    Figure Lengend Snippet: ML effect on RV replication. (A to E) Western blot and confocal immunofluorescence analyses with the indicated antibodies of RV-infected cells (MOI, 25 VFU/cell) treated with ML at 10 μM, unless otherwise indicated, or with DMSO (D) for the indicated times. Scale bars, 10 μm. TUB, tubulin. (F) Time course of viral progeny yield of OSU-infected (MOI, 25 VFU/cell) MA104 cells treated with 10 μM ML, added at 2 hpi. The data are presented as averages ± standard deviations of the results of three independent experiments. ***, P

    Article Snippet: MA104 cells (embryonic African green monkey kidney cells; ATCC CRL-2378) were grown in Dulbecco's modified Eagle's medium (DMEM) (Life Technologies) containing 10% fetal bovine serum (Life Technologies) and 50 μg/ml gentamicin (Biochrom AG).

    Techniques: Western Blot, Immunofluorescence, Infection

    ML effect on VLS. Confocal immunofluorescence assay of VLS (A and C) and Western blot analysis (B) with the indicated antibodies of MA104 cells transfected with NSP5, NSP2, VP2, and VP6, as indicated. (A) NSP5 is shown in green and NSP2 or VP2 in red. (C) NSP5 is shown in red and VP6 in green. Cells were treated for 5 h with 10 μM ML or DMSO at 18 h posttransfection. Bars, 5 μm.

    Journal: Journal of Virology

    Article Title: Identification of a Small Molecule That Compromises the Structural Integrity of Viroplasms and Rotavirus Double-Layered Particles

    doi: 10.1128/JVI.01943-17

    Figure Lengend Snippet: ML effect on VLS. Confocal immunofluorescence assay of VLS (A and C) and Western blot analysis (B) with the indicated antibodies of MA104 cells transfected with NSP5, NSP2, VP2, and VP6, as indicated. (A) NSP5 is shown in green and NSP2 or VP2 in red. (C) NSP5 is shown in red and VP6 in green. Cells were treated for 5 h with 10 μM ML or DMSO at 18 h posttransfection. Bars, 5 μm.

    Article Snippet: MA104 cells (embryonic African green monkey kidney cells; ATCC CRL-2378) were grown in Dulbecco's modified Eagle's medium (DMEM) (Life Technologies) containing 10% fetal bovine serum (Life Technologies) and 50 μg/ml gentamicin (Biochrom AG).

    Techniques: Immunofluorescence, Western Blot, Transfection

    Analysis of the interaction between proteins in MA104 cell extracts and 32 P-labeled arterivirus 3′(+)NCR RNAs. (A) Gel mobility shift assay. Radiolabeled SHFV 3′(+)NCR RNA was incubated with an S100 cytoplasmic extract from either SHFV-infected or mock-infected MA104 cells. The RPCs were resolved on a 10% non-denaturing polyacrylamide gel and visualized by autoradiography. (Lane 1) free probe; (lanes 2–5) increasing amounts of mock-infected MA104 S100 cytoplasmic extract (100, 200, 300, and 400 ng); (lanes 6–10) increasing amounts of SHFV-infected MA104 S100 cytoplasmic extract (100, 200, 300, 400, and 500 ng). The locations of the RNA–protein complex and free probe are indicated by arrows. (B) Competition gel mobility shift assay. Different amounts of non-radiolabeled competitor RNAs were incubated with an MA104 S100 cytoplasmic extract before addition of the 32 P-labeled SHFV 3′(+)NCR RNA. The RPCs were resolved on a 10% non-denaturing polyacrylamide gel and visualized by autoradiography. (Lane 1) free probe; (lane 2) no competitor; (lanes 3–6) increasing amounts of non-radiolabeled SHFV 3′(+)NCR RNA (5-, 10-, 20-, and 30-fold molar excess); (lane 7) 250-fold molar excess of yeast tRNA; (lane 8) 150-fold molar excess of WNV 3′(+)SL RNA; (lane 9) 250-fold molar excess of poly(I)–poly(C). The locations of the RNA–protein complex and free probe are indicated by arrows. (C) UV-induced cross-linking assay. MA104 S100 cytoplasmic extracts were incubated with radiolabeled SHFV 3′(+)NCR RNA and then were exposed to UV-irradiation. The unprotected RNA was digested with RNase A and the cross-linked proteins were resolved by 10% SDS–PAGE and visualized by autoradiography. (Lane 1) free probe; (lane 2) mock-infected MA104 S100 cytoplasmic extract (1 μg) and poly(I)–(C) (1 μg); (lane 3) SHFV-infected MA104 S100 cytoplasmic extract (1 μg) and poly(I)–(C) (1 μg); (lane 4) mock-infected MA104 S100 cytoplasmic extract (1 μg) and poly(I)–(C) (600 ng). Standard protein markers are indicated by lines and the positions of p56 and p42 are indicated by arrows. (D) UV-induced cross-linking assay. MA104 S100 cytoplasmic extracts and different arterivirus 32 P-labeled RNA probes were cross-linked by UV-irradiation in the presence of 600 ng of poly(I)–(C). (Lane 1) free probe; (lane 2) SHFV 3′(+)NCR RNA; (lane 3) EAV 3′(+)NCR RNA; (lane 4) PRRSV 3′(+)NCR RNA. The gels shown in lanes 1 and 2 were analyzed by autoradiography and the gel shown in lanes 3 and 4 was analyzed using the FUJI Bio Imaging Analyzer. The positions of protein standard markers are indicated by lines on the right. The positions of the p56 and p42 bands are indicated by arrows. (E) Competition gel mobility shift assay. MA104 S100 cytoplasmic extracts were incubated with different amounts of non-radiolabeled arterivirus RNAs before addition of the 32 P-labeled SHFV 3′(+)NCR RNA. (Lane 1) free probe; (lane 2) no competitor; (lanes 3–5) increasing amounts of unlabeled EAV 3′(+)NCR RNA (25-, 50-, and 75-fold molar excess); (lanes 6–8) increasing amounts of unlabeled PRRSV 3′(+)NCR RNA (25-, 50-, and 75-fold molar excess). The gels were analyzed using the FUJI Bio Imaging Analyzer. The locations of the RNA–protein complex and free probe are indicated by arrows.

    Journal: Virus Research

    Article Title: Two cellular proteins that interact with a stem loop in the simian hemorrhagic fever virus 3′(+)NCR RNA

    doi: 10.1016/j.virusres.2004.11.014

    Figure Lengend Snippet: Analysis of the interaction between proteins in MA104 cell extracts and 32 P-labeled arterivirus 3′(+)NCR RNAs. (A) Gel mobility shift assay. Radiolabeled SHFV 3′(+)NCR RNA was incubated with an S100 cytoplasmic extract from either SHFV-infected or mock-infected MA104 cells. The RPCs were resolved on a 10% non-denaturing polyacrylamide gel and visualized by autoradiography. (Lane 1) free probe; (lanes 2–5) increasing amounts of mock-infected MA104 S100 cytoplasmic extract (100, 200, 300, and 400 ng); (lanes 6–10) increasing amounts of SHFV-infected MA104 S100 cytoplasmic extract (100, 200, 300, 400, and 500 ng). The locations of the RNA–protein complex and free probe are indicated by arrows. (B) Competition gel mobility shift assay. Different amounts of non-radiolabeled competitor RNAs were incubated with an MA104 S100 cytoplasmic extract before addition of the 32 P-labeled SHFV 3′(+)NCR RNA. The RPCs were resolved on a 10% non-denaturing polyacrylamide gel and visualized by autoradiography. (Lane 1) free probe; (lane 2) no competitor; (lanes 3–6) increasing amounts of non-radiolabeled SHFV 3′(+)NCR RNA (5-, 10-, 20-, and 30-fold molar excess); (lane 7) 250-fold molar excess of yeast tRNA; (lane 8) 150-fold molar excess of WNV 3′(+)SL RNA; (lane 9) 250-fold molar excess of poly(I)–poly(C). The locations of the RNA–protein complex and free probe are indicated by arrows. (C) UV-induced cross-linking assay. MA104 S100 cytoplasmic extracts were incubated with radiolabeled SHFV 3′(+)NCR RNA and then were exposed to UV-irradiation. The unprotected RNA was digested with RNase A and the cross-linked proteins were resolved by 10% SDS–PAGE and visualized by autoradiography. (Lane 1) free probe; (lane 2) mock-infected MA104 S100 cytoplasmic extract (1 μg) and poly(I)–(C) (1 μg); (lane 3) SHFV-infected MA104 S100 cytoplasmic extract (1 μg) and poly(I)–(C) (1 μg); (lane 4) mock-infected MA104 S100 cytoplasmic extract (1 μg) and poly(I)–(C) (600 ng). Standard protein markers are indicated by lines and the positions of p56 and p42 are indicated by arrows. (D) UV-induced cross-linking assay. MA104 S100 cytoplasmic extracts and different arterivirus 32 P-labeled RNA probes were cross-linked by UV-irradiation in the presence of 600 ng of poly(I)–(C). (Lane 1) free probe; (lane 2) SHFV 3′(+)NCR RNA; (lane 3) EAV 3′(+)NCR RNA; (lane 4) PRRSV 3′(+)NCR RNA. The gels shown in lanes 1 and 2 were analyzed by autoradiography and the gel shown in lanes 3 and 4 was analyzed using the FUJI Bio Imaging Analyzer. The positions of protein standard markers are indicated by lines on the right. The positions of the p56 and p42 bands are indicated by arrows. (E) Competition gel mobility shift assay. MA104 S100 cytoplasmic extracts were incubated with different amounts of non-radiolabeled arterivirus RNAs before addition of the 32 P-labeled SHFV 3′(+)NCR RNA. (Lane 1) free probe; (lane 2) no competitor; (lanes 3–5) increasing amounts of unlabeled EAV 3′(+)NCR RNA (25-, 50-, and 75-fold molar excess); (lanes 6–8) increasing amounts of unlabeled PRRSV 3′(+)NCR RNA (25-, 50-, and 75-fold molar excess). The gels were analyzed using the FUJI Bio Imaging Analyzer. The locations of the RNA–protein complex and free probe are indicated by arrows.

    Article Snippet: Stock virus pools were prepared in MA104 cells by infecting confluent MA104 monolayers with SHFV, strain LVR 42-0/M6941 (American Type Culture Collection, passage 2), the prototype strain of SHFV, at a MOI of 0.2.

    Techniques: Labeling, Mobility Shift, Incubation, Infection, Autoradiography, Irradiation, SDS Page, Imaging

    Thermodynamically predicted secondary structures of the full length and deleted SHFV 3′(+)NCR RNAs and UV-induced cross-linking assays with these RNAs. (A) Thermodynamically predicted secondary structures of the indicated RNAs generated with Mfold ( Mathews et al., 1999 , Zuker et al., 1999 ). Δ G , the free energy values calculated for each structure. Locations of mutations within the RNAs are indicated by gray lines and arrows. (B) UV-induced cross-linking assays with an MA104 S100 cytoplasmic extract and a 32 P-labeled SHFV RNA probe in the presence of 1 μg of poly(I)–(C). (Lane 1) free probe; (lane 2) full-length SHFV 3′(+)NCR RNA; (lane 3) RNA-E; (lane 4) RNA-F; (lane 5) RNA-G. The positions of the p56 and p42 bands are indicated by arrows.

    Journal: Virus Research

    Article Title: Two cellular proteins that interact with a stem loop in the simian hemorrhagic fever virus 3′(+)NCR RNA

    doi: 10.1016/j.virusres.2004.11.014

    Figure Lengend Snippet: Thermodynamically predicted secondary structures of the full length and deleted SHFV 3′(+)NCR RNAs and UV-induced cross-linking assays with these RNAs. (A) Thermodynamically predicted secondary structures of the indicated RNAs generated with Mfold ( Mathews et al., 1999 , Zuker et al., 1999 ). Δ G , the free energy values calculated for each structure. Locations of mutations within the RNAs are indicated by gray lines and arrows. (B) UV-induced cross-linking assays with an MA104 S100 cytoplasmic extract and a 32 P-labeled SHFV RNA probe in the presence of 1 μg of poly(I)–(C). (Lane 1) free probe; (lane 2) full-length SHFV 3′(+)NCR RNA; (lane 3) RNA-E; (lane 4) RNA-F; (lane 5) RNA-G. The positions of the p56 and p42 bands are indicated by arrows.

    Article Snippet: Stock virus pools were prepared in MA104 cells by infecting confluent MA104 monolayers with SHFV, strain LVR 42-0/M6941 (American Type Culture Collection, passage 2), the prototype strain of SHFV, at a MOI of 0.2.

    Techniques: Generated, Labeling

    Effect of SLPE and LPE on rotavirus binding and infectivity of MA104 cells. ( A ) Virus (RV TLP) binding and ( B ) infectivity of MA104 cells in the presence and absence of various concentrations of SLPE, LPE, and other lipids or neoglycoconjugates were measured as described in the Experimental Section. (●), PE; ( ), LPE; ( ), SLPE; ( ), NGcGM 3 ; ( ), Sialyllactose-BSA (SL-BSA). SL-BSA concentrations are based on µM equivalent of sialic acid.

    Journal: Nutrients

    Article Title: Inhibition of Rotavirus Infectivity by a Neoglycolipid Receptor Mimetic

    doi: 10.3390/nu3020228

    Figure Lengend Snippet: Effect of SLPE and LPE on rotavirus binding and infectivity of MA104 cells. ( A ) Virus (RV TLP) binding and ( B ) infectivity of MA104 cells in the presence and absence of various concentrations of SLPE, LPE, and other lipids or neoglycoconjugates were measured as described in the Experimental Section. (●), PE; ( ), LPE; ( ), SLPE; ( ), NGcGM 3 ; ( ), Sialyllactose-BSA (SL-BSA). SL-BSA concentrations are based on µM equivalent of sialic acid.

    Article Snippet: Cells and Virus For all in vitro experiments, Group A porcine rotavirus (OSU strain (P9(7)G5)) was propagated in MA104 cells (ATCC HTB 37) and triple and double-layered virus particles isolated by gradient purification using the following modification of standard techniques [ , , ].

    Techniques: Binding Assay, Infection