baby hamster kidney bhk 21 cells  (Millipore)


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    Millipore baby hamster kidney bhk 21 cells
    Activation of RNase L during vaccinia virus infection of RoNi/7 cells requires OAS3 expression and is independent of MAVS expression. (A) WT and KO RoNi/7 cells were infected with VACVΔE3L (MOI = 1 PFU/cell), and at 6 h postinfection cells were lysed and RNA integrity was assessed on a Bioanalyzer. The positions of 18S and 28S rRNA are indicated. The data are from one representative of two independent experiments. (B) Cells were infected in triplicate with VACVΔE3L (MOI = 1 PFU/cell); at 42 h postinfection, the cells were freeze-thawed three times; and infectious virus titers were determined by plaque assay on <t>BHK-21</t> indicator cells. The viral titer data are from one of four independent experiments and expressed as means ± SDs (*, P
    Baby Hamster Kidney Bhk 21 Cells, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 90 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    baby hamster kidney bhk 21 cells - by Bioz Stars, 2020-09
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    1) Product Images from "Activation of RNase L in Egyptian Rousette Bat-Derived RoNi/7 Cells Is Dependent Primarily on OAS3 and Independent of MAVS Signaling"

    Article Title: Activation of RNase L in Egyptian Rousette Bat-Derived RoNi/7 Cells Is Dependent Primarily on OAS3 and Independent of MAVS Signaling

    Journal: mBio

    doi: 10.1128/mBio.02414-19

    Activation of RNase L during vaccinia virus infection of RoNi/7 cells requires OAS3 expression and is independent of MAVS expression. (A) WT and KO RoNi/7 cells were infected with VACVΔE3L (MOI = 1 PFU/cell), and at 6 h postinfection cells were lysed and RNA integrity was assessed on a Bioanalyzer. The positions of 18S and 28S rRNA are indicated. The data are from one representative of two independent experiments. (B) Cells were infected in triplicate with VACVΔE3L (MOI = 1 PFU/cell); at 42 h postinfection, the cells were freeze-thawed three times; and infectious virus titers were determined by plaque assay on BHK-21 indicator cells. The viral titer data are from one of four independent experiments and expressed as means ± SDs (*, P
    Figure Legend Snippet: Activation of RNase L during vaccinia virus infection of RoNi/7 cells requires OAS3 expression and is independent of MAVS expression. (A) WT and KO RoNi/7 cells were infected with VACVΔE3L (MOI = 1 PFU/cell), and at 6 h postinfection cells were lysed and RNA integrity was assessed on a Bioanalyzer. The positions of 18S and 28S rRNA are indicated. The data are from one representative of two independent experiments. (B) Cells were infected in triplicate with VACVΔE3L (MOI = 1 PFU/cell); at 42 h postinfection, the cells were freeze-thawed three times; and infectious virus titers were determined by plaque assay on BHK-21 indicator cells. The viral titer data are from one of four independent experiments and expressed as means ± SDs (*, P

    Techniques Used: Activation Assay, Infection, Expressing, Plaque Assay

    Related Articles

    Modification:

    Article Title: Genome-wide profiling of host-encoded circular RNAs highlights their potential role during the Japanese encephalitis virus-induced neuroinflammatory response
    Article Snippet: .. Cell and virusMouse microglial cells, baby hamster kidney (BHK-21) cells, and human embryonic kidney (HEK293T) cells were cultured and maintained in Dulbecco modified Eagle medium (DMEM; Sigma) supplement with 10% fetal bovine serum (FBS; Gibco), 100 U/ml penicillin, and 100 mg/ml streptomycin sulfate at 37 °C in 5% CO2 atmosphere. .. The circRNA knockdown BV2 cells were engineered by the CRISPR-Cas9 system.

    Article Title: Expression of the E3L Gene of Vaccinia Virus in Transgenic Mice Decreases Host Resistance to Vaccinia Virus and Leishmania major Infections ▿
    Article Snippet: .. Baby hamster kidney BHK-21 cells were grown in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal calf serum (FCS), l -glutamine (2 mM; Sigma), penicillin (100 U/ml; Sigma), streptomycin (0.1 mg/ml; Sigma), gentamicin (5 μg/ml; Sigma), and amphotericin B (Fungizone) (0.5 μg/ml; Sigma). ..

    Cell Culture:

    Article Title: Activation of RNase L is dependent on OAS3 expression during infection with diverse human viruses
    Article Snippet: .. Baby hamster kidney (BHK-21) cells were cultured in DMEM supplemented with 10% (vol/vol) FBS, 5% (vol/vol) tryptose phosphate broth (Sigma-Aldrich), 100 U/mL penicillin, and 100 μg/mL streptomycin. .. Human HEK 293T cells were cultured in DMEM supplemented with 10% (vol/vol) FBS and 1 mM sodium pyruvate.

    Article Title: Genome-wide profiling of host-encoded circular RNAs highlights their potential role during the Japanese encephalitis virus-induced neuroinflammatory response
    Article Snippet: .. Cell and virusMouse microglial cells, baby hamster kidney (BHK-21) cells, and human embryonic kidney (HEK293T) cells were cultured and maintained in Dulbecco modified Eagle medium (DMEM; Sigma) supplement with 10% fetal bovine serum (FBS; Gibco), 100 U/ml penicillin, and 100 mg/ml streptomycin sulfate at 37 °C in 5% CO2 atmosphere. .. The circRNA knockdown BV2 cells were engineered by the CRISPR-Cas9 system.

    Article Title: Activation of RNase L in Egyptian Rousette Bat-Derived RoNi/7 Cells Is Dependent Primarily on OAS3 and Independent of MAVS Signaling
    Article Snippet: .. Baby hamster kidney (BHK-21) cells were cultured in DMEM supplemented with 10% FBS, 5% tryptose phosphate broth (Sigma-Aldrich), 100 U/ml penicillin, and 100 mg/ml streptomycin. .. Human HEK 293T cells were cultured in DMEM supplemented with 10% FBS and 1 mM sodium pyruvate.

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    Millipore bhk 21 cells
    Knocking-down the expression of GRP78 does not affect the replication of JE viral RNA . <t>BHK-21</t> cells were transfected with siRNA against GRP78 or an irrelevant siRNA (scramble siRNA) for 48 hours. The expression level of GRP78 was measured by Western blot analysis using polyclonal antibody specific to GRP78. Transfected cells were then infected with JEV at an MOI of 1. At 24 hour post-infection, the cell lysates were collected to measure the JEV replication using antibodies specific to NS1 and/or NS5. Two independent replicates are shown for the scramble and GRP78 siRNA conditions
    Bhk 21 Cells, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Knocking-down the expression of GRP78 does not affect the replication of JE viral RNA . BHK-21 cells were transfected with siRNA against GRP78 or an irrelevant siRNA (scramble siRNA) for 48 hours. The expression level of GRP78 was measured by Western blot analysis using polyclonal antibody specific to GRP78. Transfected cells were then infected with JEV at an MOI of 1. At 24 hour post-infection, the cell lysates were collected to measure the JEV replication using antibodies specific to NS1 and/or NS5. Two independent replicates are shown for the scramble and GRP78 siRNA conditions

    Journal: Virology Journal

    Article Title: Japanese encephalitis virus co-opts the ER-stress response protein GRP78 for viral infectivity

    doi: 10.1186/1743-422X-8-128

    Figure Lengend Snippet: Knocking-down the expression of GRP78 does not affect the replication of JE viral RNA . BHK-21 cells were transfected with siRNA against GRP78 or an irrelevant siRNA (scramble siRNA) for 48 hours. The expression level of GRP78 was measured by Western blot analysis using polyclonal antibody specific to GRP78. Transfected cells were then infected with JEV at an MOI of 1. At 24 hour post-infection, the cell lysates were collected to measure the JEV replication using antibodies specific to NS1 and/or NS5. Two independent replicates are shown for the scramble and GRP78 siRNA conditions

    Article Snippet: Sucrose density gradient analysis The secretion medium of infected BHK-21 cells (2 days post-infection at MOI of 10) was centrifuged at 6,000 rpm for 20 minutes in 4°C to remove cell debris, and was concentrated with a concentration tube (Millipore, MA, USA) at 6,000 rpm for 20 minutes.

    Techniques: Expressing, Transfection, Western Blot, Infection

    Proteins identified by LC-MS in the secretion medium of JEV-infected BHK-21 cells . Cell extracts were collected at three days after mock- and JEV-infection and subjected to one dimension SDS-PAGE analysis. A total amount of 10 μg of secreted proteins from mock- and JEV-infected BHK-21 cells was loaded per lane. The gel was stained with Silver nitrate. The identified proteins are shown as Table 1.

    Journal: Virology Journal

    Article Title: Japanese encephalitis virus co-opts the ER-stress response protein GRP78 for viral infectivity

    doi: 10.1186/1743-422X-8-128

    Figure Lengend Snippet: Proteins identified by LC-MS in the secretion medium of JEV-infected BHK-21 cells . Cell extracts were collected at three days after mock- and JEV-infection and subjected to one dimension SDS-PAGE analysis. A total amount of 10 μg of secreted proteins from mock- and JEV-infected BHK-21 cells was loaded per lane. The gel was stained with Silver nitrate. The identified proteins are shown as Table 1.

    Article Snippet: Sucrose density gradient analysis The secretion medium of infected BHK-21 cells (2 days post-infection at MOI of 10) was centrifuged at 6,000 rpm for 20 minutes in 4°C to remove cell debris, and was concentrated with a concentration tube (Millipore, MA, USA) at 6,000 rpm for 20 minutes.

    Techniques: Liquid Chromatography with Mass Spectroscopy, Infection, SDS Page, Staining

    GRP78 is released into the media upon JEV infection and partially co-fractionates with the JE virion . A) Verification of GRP78 in the secretome from JEV-induced BHK-21 cells. Cell lysates or secretion medium were collected 3 days post-infection followed by SDS-PAGE for protein separation. The GRP78 was detected by anti-GRP78 specific antibody. Two independent replicates are shown for the mock- and JEV-infected conditions. B) Sucrose density gradient fraction of JE virion and GPR78. A volume of 40 μL of sample from each fraction was analyzed on SDS-PAGE followed by the detection of anti-JEV E protein and anti-GRP78 by Western blotting.

    Journal: Virology Journal

    Article Title: Japanese encephalitis virus co-opts the ER-stress response protein GRP78 for viral infectivity

    doi: 10.1186/1743-422X-8-128

    Figure Lengend Snippet: GRP78 is released into the media upon JEV infection and partially co-fractionates with the JE virion . A) Verification of GRP78 in the secretome from JEV-induced BHK-21 cells. Cell lysates or secretion medium were collected 3 days post-infection followed by SDS-PAGE for protein separation. The GRP78 was detected by anti-GRP78 specific antibody. Two independent replicates are shown for the mock- and JEV-infected conditions. B) Sucrose density gradient fraction of JE virion and GPR78. A volume of 40 μL of sample from each fraction was analyzed on SDS-PAGE followed by the detection of anti-JEV E protein and anti-GRP78 by Western blotting.

    Article Snippet: Sucrose density gradient analysis The secretion medium of infected BHK-21 cells (2 days post-infection at MOI of 10) was centrifuged at 6,000 rpm for 20 minutes in 4°C to remove cell debris, and was concentrated with a concentration tube (Millipore, MA, USA) at 6,000 rpm for 20 minutes.

    Techniques: Infection, SDS Page, Western Blot

    The co-localization of GRP78 and JEV E protein in JEV-infected BHK-21 cells . Mock- or JEV-infected BHK-21 cells were harvested at 3 days post-infection and prepared for immunofluorescence analysis stained with antibodies that detect GRP78 (green; b, f, j) and JEV-E protein (red; a, e, i).

    Journal: Virology Journal

    Article Title: Japanese encephalitis virus co-opts the ER-stress response protein GRP78 for viral infectivity

    doi: 10.1186/1743-422X-8-128

    Figure Lengend Snippet: The co-localization of GRP78 and JEV E protein in JEV-infected BHK-21 cells . Mock- or JEV-infected BHK-21 cells were harvested at 3 days post-infection and prepared for immunofluorescence analysis stained with antibodies that detect GRP78 (green; b, f, j) and JEV-E protein (red; a, e, i).

    Article Snippet: Sucrose density gradient analysis The secretion medium of infected BHK-21 cells (2 days post-infection at MOI of 10) was centrifuged at 6,000 rpm for 20 minutes in 4°C to remove cell debris, and was concentrated with a concentration tube (Millipore, MA, USA) at 6,000 rpm for 20 minutes.

    Techniques: Infection, Immunofluorescence, Staining

    Knocking-down the expression of GRP78 by siRNA decreases the yield of infectious JE virus production . BHK-21 cells were transfected with siRNA against GRP78 or an irrelevant siRNA (negative control). The down-regulation of GRP78 was measured by Western blot analysis using antibody specific to GRP78 as shown in Figure 5. A) Transfected cell lysates were then infected with JEV at an MOI of 1. At 24 hours post-infection, the supernatants were collected to measure the amount of JE viral RNA production by RT real-time PCR as described in Material and Methods. The virus yield is expressed as a percentage of the yield obtained from cells transfected with irrelevant siRNA. B) Plaque formation by JE virus-particle collected from JEV-infected scramble siRNA treated cells or cells treated with siRNA against GRP78. C) Quantitative measurement of viral progeny produced from JEV-infected scramble siRNA treated cells or cells treated with siRNA against GRP78. The virus titer is defined as plaque-forming unit (PFU) per mL. Results are derived from three independent experiments.

    Journal: Virology Journal

    Article Title: Japanese encephalitis virus co-opts the ER-stress response protein GRP78 for viral infectivity

    doi: 10.1186/1743-422X-8-128

    Figure Lengend Snippet: Knocking-down the expression of GRP78 by siRNA decreases the yield of infectious JE virus production . BHK-21 cells were transfected with siRNA against GRP78 or an irrelevant siRNA (negative control). The down-regulation of GRP78 was measured by Western blot analysis using antibody specific to GRP78 as shown in Figure 5. A) Transfected cell lysates were then infected with JEV at an MOI of 1. At 24 hours post-infection, the supernatants were collected to measure the amount of JE viral RNA production by RT real-time PCR as described in Material and Methods. The virus yield is expressed as a percentage of the yield obtained from cells transfected with irrelevant siRNA. B) Plaque formation by JE virus-particle collected from JEV-infected scramble siRNA treated cells or cells treated with siRNA against GRP78. C) Quantitative measurement of viral progeny produced from JEV-infected scramble siRNA treated cells or cells treated with siRNA against GRP78. The virus titer is defined as plaque-forming unit (PFU) per mL. Results are derived from three independent experiments.

    Article Snippet: Sucrose density gradient analysis The secretion medium of infected BHK-21 cells (2 days post-infection at MOI of 10) was centrifuged at 6,000 rpm for 20 minutes in 4°C to remove cell debris, and was concentrated with a concentration tube (Millipore, MA, USA) at 6,000 rpm for 20 minutes.

    Techniques: Expressing, Transfection, Negative Control, Western Blot, Infection, Real-time Polymerase Chain Reaction, Produced, Derivative Assay

    An assay for the collection of secreted proteins from JEV-infected BHK-21 cells . A) A flowchart outlining the protocol for the identification of JE virus replication from intracellular lysates and secreted proteins from the JEV-infected cells. B) Effect of serum deprivation on JEV replication. Cells were incubated with medium supplemented with 2% FBS or serum-free medium for 24 hours and viral proteins, NS1 and NS5, were assessed by Western blot analysis using anti-NS5/anti-NS1 polyclonal antibodies. Extracts from mock-infected cells serve as a negative control. Two independent replicates are shown for the serum-free and 2% serum conditions. C) Effect of serum deprivation on cell viability. β-actin was not detected in the secreted medium of mock- and JEV-infected BHK-21 cells. Three independent replicates are shown for the mock- and JEV-infected conditions.

    Journal: Virology Journal

    Article Title: Japanese encephalitis virus co-opts the ER-stress response protein GRP78 for viral infectivity

    doi: 10.1186/1743-422X-8-128

    Figure Lengend Snippet: An assay for the collection of secreted proteins from JEV-infected BHK-21 cells . A) A flowchart outlining the protocol for the identification of JE virus replication from intracellular lysates and secreted proteins from the JEV-infected cells. B) Effect of serum deprivation on JEV replication. Cells were incubated with medium supplemented with 2% FBS or serum-free medium for 24 hours and viral proteins, NS1 and NS5, were assessed by Western blot analysis using anti-NS5/anti-NS1 polyclonal antibodies. Extracts from mock-infected cells serve as a negative control. Two independent replicates are shown for the serum-free and 2% serum conditions. C) Effect of serum deprivation on cell viability. β-actin was not detected in the secreted medium of mock- and JEV-infected BHK-21 cells. Three independent replicates are shown for the mock- and JEV-infected conditions.

    Article Snippet: Sucrose density gradient analysis The secretion medium of infected BHK-21 cells (2 days post-infection at MOI of 10) was centrifuged at 6,000 rpm for 20 minutes in 4°C to remove cell debris, and was concentrated with a concentration tube (Millipore, MA, USA) at 6,000 rpm for 20 minutes.

    Techniques: Infection, Incubation, Western Blot, Negative Control

    Replication in mosquito cells. (a) Growth kinetics. A. albopictus C6/36 cells were infected with rSBV or rSBVdelNSs at an m.o.i. of 3 and incubated at 28 °C. The amount of virus released into the supernatant at different times p.i. was determined by plaque assay in BHK-21 cells. (b) N production. Cells were infected as above and lysates were prepared at different times p.i. Proteins were separated by SDS-PAGE and transferred to nitrocellulose membrane. Accumulation of viral N protein was detected by reaction with anti-N antibody, and detection of tubulin (tub) was used as a loading control. m, Mock-infected cells.

    Journal: The Journal of General Virology

    Article Title: Establishment of a reverse genetics system for Schmallenberg virus, a newly emerged orthobunyavirus in Europe

    doi: 10.1099/vir.0.049981-0

    Figure Lengend Snippet: Replication in mosquito cells. (a) Growth kinetics. A. albopictus C6/36 cells were infected with rSBV or rSBVdelNSs at an m.o.i. of 3 and incubated at 28 °C. The amount of virus released into the supernatant at different times p.i. was determined by plaque assay in BHK-21 cells. (b) N production. Cells were infected as above and lysates were prepared at different times p.i. Proteins were separated by SDS-PAGE and transferred to nitrocellulose membrane. Accumulation of viral N protein was detected by reaction with anti-N antibody, and detection of tubulin (tub) was used as a loading control. m, Mock-infected cells.

    Article Snippet: SBV was grown in BHK-21 cells at 37 °C until a marked CPE was noted, and 1.75 ml clarified supernatant medium was concentrated approximately tenfold using an Amicon Ultra-15 centrifugal filter device (Millipore).

    Techniques: Infection, Incubation, Plaque Assay, SDS Page

    Plaque phenotypes of SBV in different cell lines. Infected cells were fixed after 3 days incubation at 37 °C and stained with crystal violet. (a–d) Wild-type (authentic) SBV in BHK-21 (a), BSR-T7/5 (b), Vero E6 (c) and CPT-Tert (d) cells; (e) rSBV in BHK-21 cells; (f) rSBVdelNSs in BHK-21 cells.

    Journal: The Journal of General Virology

    Article Title: Establishment of a reverse genetics system for Schmallenberg virus, a newly emerged orthobunyavirus in Europe

    doi: 10.1099/vir.0.049981-0

    Figure Lengend Snippet: Plaque phenotypes of SBV in different cell lines. Infected cells were fixed after 3 days incubation at 37 °C and stained with crystal violet. (a–d) Wild-type (authentic) SBV in BHK-21 (a), BSR-T7/5 (b), Vero E6 (c) and CPT-Tert (d) cells; (e) rSBV in BHK-21 cells; (f) rSBVdelNSs in BHK-21 cells.

    Article Snippet: SBV was grown in BHK-21 cells at 37 °C until a marked CPE was noted, and 1.75 ml clarified supernatant medium was concentrated approximately tenfold using an Amicon Ultra-15 centrifugal filter device (Millipore).

    Techniques: Infection, Incubation, Staining, Cycling Probe Technology

    Viral protein synthesis in mammalian cells. (a) N production. BHK-21, CPT-Tert or A549 cells were infected with wtSBV, rSBV or rSBVdelNSs at an m.o.i. of 3 and incubated at 37 °C. Cell lysates were prepared at different times after infection, and proteins were separated by SDS-PAGE and transferred to nitrocellulose membrane. Accumulation of viral N was detected by reaction with anti-N antibody, and detection of tubulin (tub) was used as a loading control. (b) Host-cell protein shut-off. BHK-21 cells were infected with wtSBV, rSBV or rSBVdelNSs at an m.o.i. of 3 and incubated at 37 °C. At different times p.i., the cells were labelled for 1 h with [ 35 S]methionine and cell extracts were fractionated by SDS-PAGE. The dried gel was exposed to X-ray film. The positions of viral Gc and N are indicated. m, Mock-infected cells.

    Journal: The Journal of General Virology

    Article Title: Establishment of a reverse genetics system for Schmallenberg virus, a newly emerged orthobunyavirus in Europe

    doi: 10.1099/vir.0.049981-0

    Figure Lengend Snippet: Viral protein synthesis in mammalian cells. (a) N production. BHK-21, CPT-Tert or A549 cells were infected with wtSBV, rSBV or rSBVdelNSs at an m.o.i. of 3 and incubated at 37 °C. Cell lysates were prepared at different times after infection, and proteins were separated by SDS-PAGE and transferred to nitrocellulose membrane. Accumulation of viral N was detected by reaction with anti-N antibody, and detection of tubulin (tub) was used as a loading control. (b) Host-cell protein shut-off. BHK-21 cells were infected with wtSBV, rSBV or rSBVdelNSs at an m.o.i. of 3 and incubated at 37 °C. At different times p.i., the cells were labelled for 1 h with [ 35 S]methionine and cell extracts were fractionated by SDS-PAGE. The dried gel was exposed to X-ray film. The positions of viral Gc and N are indicated. m, Mock-infected cells.

    Article Snippet: SBV was grown in BHK-21 cells at 37 °C until a marked CPE was noted, and 1.75 ml clarified supernatant medium was concentrated approximately tenfold using an Amicon Ultra-15 centrifugal filter device (Millipore).

    Techniques: Cycling Probe Technology, Infection, Incubation, SDS Page

    Virus growth curves. BHK-21, CPT-Tert or A549 cells were infected with wtSBV, rSBV or rSBVdelNSs at an m.o.i. of 3 and incubated at 37 °C. The titre of virus released into the supernatant at different times post-infection (p.i.) was determined by plaque assay in BHK-21 cells.

    Journal: The Journal of General Virology

    Article Title: Establishment of a reverse genetics system for Schmallenberg virus, a newly emerged orthobunyavirus in Europe

    doi: 10.1099/vir.0.049981-0

    Figure Lengend Snippet: Virus growth curves. BHK-21, CPT-Tert or A549 cells were infected with wtSBV, rSBV or rSBVdelNSs at an m.o.i. of 3 and incubated at 37 °C. The titre of virus released into the supernatant at different times post-infection (p.i.) was determined by plaque assay in BHK-21 cells.

    Article Snippet: SBV was grown in BHK-21 cells at 37 °C until a marked CPE was noted, and 1.75 ml clarified supernatant medium was concentrated approximately tenfold using an Amicon Ultra-15 centrifugal filter device (Millipore).

    Techniques: Cycling Probe Technology, Infection, Incubation, Plaque Assay

    Expression and subcellular localization of HA and NA in HA-NA co-transfected BHK-21 cells by immunofluorescence microscopy assay. Cell nuclei were labeled with DAPI (blue), and HAs/NAs were labeled with anti-HA (clone HA-7) and anti-N2 (H6N2) primary antibodies and with corresponding fluorophore-conjugated antibodies (false-colored with HA in green and NA in red). Top three rows: permeabilized BHK-21 cells imaged using 100 × objective. Bottom three rows: non-permeabilized BHK-21 cells using 100 × objective and with identical exposure time. For each panel, cells were co-transfected with pCAGGS-H3(X-31) and with either (from left to right): pCAGGS-empty (No NA), pCAGGS-N2 (X-31), pCAGGS-N2(Japan) (Japan N2 NA), and pCAGGS-N2(MS96) (MS96 N2 NA). Scale bar represents 10 μm.

    Journal: Scientific Reports

    Article Title: Viral fusion efficacy of specific H3N2 influenza virus reassortant combinations at single-particle level

    doi: 10.1038/srep35537

    Figure Lengend Snippet: Expression and subcellular localization of HA and NA in HA-NA co-transfected BHK-21 cells by immunofluorescence microscopy assay. Cell nuclei were labeled with DAPI (blue), and HAs/NAs were labeled with anti-HA (clone HA-7) and anti-N2 (H6N2) primary antibodies and with corresponding fluorophore-conjugated antibodies (false-colored with HA in green and NA in red). Top three rows: permeabilized BHK-21 cells imaged using 100 × objective. Bottom three rows: non-permeabilized BHK-21 cells using 100 × objective and with identical exposure time. For each panel, cells were co-transfected with pCAGGS-H3(X-31) and with either (from left to right): pCAGGS-empty (No NA), pCAGGS-N2 (X-31), pCAGGS-N2(Japan) (Japan N2 NA), and pCAGGS-N2(MS96) (MS96 N2 NA). Scale bar represents 10 μm.

    Article Snippet: Immunofluorescence assay 5 × 105 cells BHK-21 cells were seeded in 8-well glass slides (Millipore).

    Techniques: Expressing, Transfection, Immunofluorescence, Microscopy, Labeling

    Use of r3LCMV CAT/FLuc to identify antiarenaviral compounds. ( A ) BHK-21 cells (96-well plate) were infected (moi = 0.1) with either LCMV WT or r3LCMV CAT/FLuc and at different time points, virus titers and FLuc activity were determined. ( B ) The correlation

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Generation of recombinant lymphocytic choriomeningitis viruses with trisegmented genomes stably expressing two additional genes of interest

    doi: 10.1073/pnas.0900088106

    Figure Lengend Snippet: Use of r3LCMV CAT/FLuc to identify antiarenaviral compounds. ( A ) BHK-21 cells (96-well plate) were infected (moi = 0.1) with either LCMV WT or r3LCMV CAT/FLuc and at different time points, virus titers and FLuc activity were determined. ( B ) The correlation

    Article Snippet: Supernatants from infected BHK-21 were filtrated by using Amicon Ultra-15 100K columns (Millipore) for 10 min at 1,500 × g to concentrate virus particles.

    Techniques: Infection, Activity Assay

    Growth properties and foreign gene expression of r3LCMV GFP/CAT in cultured cells. BHK-21 cells were infected (moi of 0.1) either with LCMV WT or r3LCMV GFP/CAT. At the indicated time points, virus titers in supernatants were determined ( A ), and cells

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Generation of recombinant lymphocytic choriomeningitis viruses with trisegmented genomes stably expressing two additional genes of interest

    doi: 10.1073/pnas.0900088106

    Figure Lengend Snippet: Growth properties and foreign gene expression of r3LCMV GFP/CAT in cultured cells. BHK-21 cells were infected (moi of 0.1) either with LCMV WT or r3LCMV GFP/CAT. At the indicated time points, virus titers in supernatants were determined ( A ), and cells

    Article Snippet: Supernatants from infected BHK-21 were filtrated by using Amicon Ultra-15 100K columns (Millipore) for 10 min at 1,500 × g to concentrate virus particles.

    Techniques: Expressing, Cell Culture, Infection

    Phenotypic stability of r3LCMV GFP/CAT during serial passages. LCMV WT and r3LCMV GFP/CAT were serially passed on BHK-21 cells. At each passage, virus titers in supernatants were determined ( A ), and r3LCMV GFP/CAT-infected cells were collected for CAT

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Generation of recombinant lymphocytic choriomeningitis viruses with trisegmented genomes stably expressing two additional genes of interest

    doi: 10.1073/pnas.0900088106

    Figure Lengend Snippet: Phenotypic stability of r3LCMV GFP/CAT during serial passages. LCMV WT and r3LCMV GFP/CAT were serially passed on BHK-21 cells. At each passage, virus titers in supernatants were determined ( A ), and r3LCMV GFP/CAT-infected cells were collected for CAT

    Article Snippet: Supernatants from infected BHK-21 were filtrated by using Amicon Ultra-15 100K columns (Millipore) for 10 min at 1,500 × g to concentrate virus particles.

    Techniques: Infection

    Growth properties of r3LCMV in mice. ( A ) Growth properties in cultured cells. BHK-21 (IFN−) and L929 (IFN+) cells were infected with the indicated viruses (moi = 0.01), and at the indicated hour p.i., virus titers in supernatants were determined.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Generation of recombinant lymphocytic choriomeningitis viruses with trisegmented genomes stably expressing two additional genes of interest

    doi: 10.1073/pnas.0900088106

    Figure Lengend Snippet: Growth properties of r3LCMV in mice. ( A ) Growth properties in cultured cells. BHK-21 (IFN−) and L929 (IFN+) cells were infected with the indicated viruses (moi = 0.01), and at the indicated hour p.i., virus titers in supernatants were determined.

    Article Snippet: Supernatants from infected BHK-21 were filtrated by using Amicon Ultra-15 100K columns (Millipore) for 10 min at 1,500 × g to concentrate virus particles.

    Techniques: Mouse Assay, Cell Culture, Infection