baby hamster kidney cells bhk 21  (ATCC)


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  • 99
    Name:
    BHK 21 C 13
    Description:

    Catalog Number:
    ccl-10
    Price:
    None
    Applications:
    The World Organization for Animal Health (OIE) uses these cells for routine diagnosis of rabies.  This cell line has been used as a host for transformation with expression vectors containing selectable and amplifiable marker DNAs (e.g., Factor VIII, see ATCC CRL-8544).
    Host:
    Mesocricetus auratus, hamster, Syrian golden
    Cell Type:
    fibroblast
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    Structured Review

    ATCC baby hamster kidney cells bhk 21
    Generation and identification of VSV-846 vaccine. (A) The schematic of pVSV-XN2-846 plasmid. The triple-antigen fusion TFP846 (Rv3615c-Mtb10.4-Rv2660c) was amplified from p846 plasmid, and the triple-antigen fusion gene was then cloned into the fifth position of the pVSV-XN2 plasmid after cleavage with XhoI and NheI. (B) The microscope images of <t>BHK-21</t> cells infected with VSV-846 or non-infected mock control 6 h post-infection (200×). (C) The triple-antigen fusion TFP846 was expressed in VSV-846 infected cells. The blot was probed with anti-flag antibody. Control: Normal cells without infection.

    https://www.bioz.com/result/baby hamster kidney cells bhk 21/product/ATCC
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    baby hamster kidney cells bhk 21 - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Vesicular Stomatitis Virus-Vectored Multi-Antigen Tuberculosis Vaccine Limits Bacterial Proliferation in Mice following a Single Intranasal Dose"

    Article Title: Vesicular Stomatitis Virus-Vectored Multi-Antigen Tuberculosis Vaccine Limits Bacterial Proliferation in Mice following a Single Intranasal Dose

    Journal: Frontiers in Cellular and Infection Microbiology

    doi: 10.3389/fcimb.2017.00034

    Generation and identification of VSV-846 vaccine. (A) The schematic of pVSV-XN2-846 plasmid. The triple-antigen fusion TFP846 (Rv3615c-Mtb10.4-Rv2660c) was amplified from p846 plasmid, and the triple-antigen fusion gene was then cloned into the fifth position of the pVSV-XN2 plasmid after cleavage with XhoI and NheI. (B) The microscope images of BHK-21 cells infected with VSV-846 or non-infected mock control 6 h post-infection (200×). (C) The triple-antigen fusion TFP846 was expressed in VSV-846 infected cells. The blot was probed with anti-flag antibody. Control: Normal cells without infection.
    Figure Legend Snippet: Generation and identification of VSV-846 vaccine. (A) The schematic of pVSV-XN2-846 plasmid. The triple-antigen fusion TFP846 (Rv3615c-Mtb10.4-Rv2660c) was amplified from p846 plasmid, and the triple-antigen fusion gene was then cloned into the fifth position of the pVSV-XN2 plasmid after cleavage with XhoI and NheI. (B) The microscope images of BHK-21 cells infected with VSV-846 or non-infected mock control 6 h post-infection (200×). (C) The triple-antigen fusion TFP846 was expressed in VSV-846 infected cells. The blot was probed with anti-flag antibody. Control: Normal cells without infection.

    Techniques Used: Plasmid Preparation, Amplification, Clone Assay, Microscopy, Infection

    2) Product Images from "Induction of a Protective Response in Mice by the Dengue Virus NS3 Protein Using DNA Vaccines"

    Article Title: Induction of a Protective Response in Mice by the Dengue Virus NS3 Protein Using DNA Vaccines

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0025685

    Immunofluorescence of BHK-21 cells transfected with recombinant plasmids based on NS3 protein or its functional domains: pcTPANS3 (a), pcTPANS3H (b), pcTPANS3P (c), pcNS3 (d), pcNS3H (e) and pcNS3P (e). Cells were treated with a mouse polyclonal antibody against the DENV2 NS3 protein. Magnification 400x.
    Figure Legend Snippet: Immunofluorescence of BHK-21 cells transfected with recombinant plasmids based on NS3 protein or its functional domains: pcTPANS3 (a), pcTPANS3H (b), pcTPANS3P (c), pcNS3 (d), pcNS3H (e) and pcNS3P (e). Cells were treated with a mouse polyclonal antibody against the DENV2 NS3 protein. Magnification 400x.

    Techniques Used: Immunofluorescence, Transfection, Recombinant, Functional Assay

    Western blots of whole-cell extract (a) and culture supernatants (b) of BHK-21 cells transfected with the different DNA vaccines. The recombinant proteins were detected in SDS-PAGE, using a mouse polyclonal antibody against the DENV2 NS3, harvested from transfections with pcTPA (lane 1), pcTPANS3P (lane 2), pcTPANS3H (lane 3), pcTPANS3 (lane 4), pcNS3P (lane 5), pcNS3H (lane 6) and pcNS3 (lane 7). Arrows indicate bands corresponding to the protease, helicase and full-length NS3 recombinant proteins.
    Figure Legend Snippet: Western blots of whole-cell extract (a) and culture supernatants (b) of BHK-21 cells transfected with the different DNA vaccines. The recombinant proteins were detected in SDS-PAGE, using a mouse polyclonal antibody against the DENV2 NS3, harvested from transfections with pcTPA (lane 1), pcTPANS3P (lane 2), pcTPANS3H (lane 3), pcTPANS3 (lane 4), pcNS3P (lane 5), pcNS3H (lane 6) and pcNS3 (lane 7). Arrows indicate bands corresponding to the protease, helicase and full-length NS3 recombinant proteins.

    Techniques Used: Western Blot, Transfection, Recombinant, SDS Page

    3) Product Images from "Foreign Gene Transfer in Termite Cells Using a Recombinant Vesicular Stomatitis Virus"

    Article Title: Foreign Gene Transfer in Termite Cells Using a Recombinant Vesicular Stomatitis Virus

    Journal: Journal of Insect Science

    doi: 10.1673/031.008.5201

    De novo replication of VSV-GFP in Coptotermes formosanus embryonic primary cultures. Supernatant of VSV-GFP infected termite cells collected at 1, 24, 48, 72, and 96 h post-infection was titered on BHK-21 cells in ten-fold triplicate serial dilutions. Titers were scored in pfu/ml as the highest dilution that yielded at least1 pfu.
    Figure Legend Snippet: De novo replication of VSV-GFP in Coptotermes formosanus embryonic primary cultures. Supernatant of VSV-GFP infected termite cells collected at 1, 24, 48, 72, and 96 h post-infection was titered on BHK-21 cells in ten-fold triplicate serial dilutions. Titers were scored in pfu/ml as the highest dilution that yielded at least1 pfu.

    Techniques Used: Infection

    Anti-GFP immunoblots of VSV-GFP infected embryonic termite cells. Lane 1, VSV-GFP infected Coptotermes formosanus cells at 96 h post-infection. Lane 2, uninfected C. formosanus cells. Lane 3, VSV-GFP infected Reticulitermes flavipes cells at 48 h post-infection. Lane 4, uninfected R. flavipes cells. Lane 5, VSV-GFP infected BHK-21 cells. Lane 6, uninfected BHK-21 cells. Gels were run under the same conditions. Molecular size markers (kDa) are shown on the right. The GFP band is marked by an arrow on the left.
    Figure Legend Snippet: Anti-GFP immunoblots of VSV-GFP infected embryonic termite cells. Lane 1, VSV-GFP infected Coptotermes formosanus cells at 96 h post-infection. Lane 2, uninfected C. formosanus cells. Lane 3, VSV-GFP infected Reticulitermes flavipes cells at 48 h post-infection. Lane 4, uninfected R. flavipes cells. Lane 5, VSV-GFP infected BHK-21 cells. Lane 6, uninfected BHK-21 cells. Gels were run under the same conditions. Molecular size markers (kDa) are shown on the right. The GFP band is marked by an arrow on the left.

    Techniques Used: Western Blot, Infection

    4) Product Images from "Rapid one-step construction of a Middle East Respiratory Syndrome (MERS-CoV) infectious clone system by homologous recombination"

    Article Title: Rapid one-step construction of a Middle East Respiratory Syndrome (MERS-CoV) infectious clone system by homologous recombination

    Journal: Journal of Virological Methods

    doi: 10.1016/j.jviromet.2016.07.022

    Supernatant from the BHK-21 and Vero cell overlay transfected with MERS-CoV/pYES1L plasmid was blind passaged three times to infect a fresh culture of Vero cells. These Vero cells were incubated for 3 days and observed for CPE. CPE was noted in the cells infected with the blind passage supernatant and not the negative control, indicating rescue of infectious virus (rMERS-CoV).
    Figure Legend Snippet: Supernatant from the BHK-21 and Vero cell overlay transfected with MERS-CoV/pYES1L plasmid was blind passaged three times to infect a fresh culture of Vero cells. These Vero cells were incubated for 3 days and observed for CPE. CPE was noted in the cells infected with the blind passage supernatant and not the negative control, indicating rescue of infectious virus (rMERS-CoV).

    Techniques Used: Transfection, Plasmid Preparation, Incubation, Infection, Negative Control

    Related Articles

    Electroporation:

    Article Title: Mutating Conserved Cysteines in the Alphavirus E2 Glycoprotein Causes Virus-Specific Assembly Defects
    Article Snippet: .. Electroporation of BHK-21 cells was performed as previously described ( ). ..

    In Vitro:

    Article Title: Mutating Conserved Cysteines in the Alphavirus E2 Glycoprotein Causes Virus-Specific Assembly Defects
    Article Snippet: .. BHK-21 cells were electroporated with in vitro -transcribed wild-type or mutant viral RNA, resuspended in BHK cell medium, and incubated at 37°C in the presence of 5% CO2 . ..

    Mutagenesis:

    Article Title: Mutating Conserved Cysteines in the Alphavirus E2 Glycoprotein Causes Virus-Specific Assembly Defects
    Article Snippet: .. BHK-21 cells were electroporated with in vitro -transcribed wild-type or mutant viral RNA, resuspended in BHK cell medium, and incubated at 37°C in the presence of 5% CO2 . ..

    Article Title: Murine Gammaherpesvirus 68 Lacking gp150 Shows Defective Virion Release but Establishes Normal Latency In Vivo
    Article Snippet: .. A similar phenotype was observed with the independent mutant (M7− STOP) and with growth in BHK-21 cells, as well as MEFs (Fig. ). ..

    Cell Culture:

    Article Title: A Novel Dengue Virus Inhibitor, BP13944, Discovered by High-Throughput Screening with Dengue Virus Replicon Cells Selects for Resistance in the Viral NS2B/NS3 Protease
    Article Snippet: .. BHK-21 cells (ATCC CCL-10) were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 4.5 g/liter glucose and 5% fetal bovine serum (FBS). .. The cells were cultured at 37°C in a 5% CO2 incubator.

    Incubation:

    Article Title: Mutating Conserved Cysteines in the Alphavirus E2 Glycoprotein Causes Virus-Specific Assembly Defects
    Article Snippet: .. BHK-21 cells were electroporated with in vitro -transcribed wild-type or mutant viral RNA, resuspended in BHK cell medium, and incubated at 37°C in the presence of 5% CO2 . ..

    other:

    Article Title: Mutating Conserved Cysteines in the Alphavirus E2 Glycoprotein Causes Virus-Specific Assembly Defects
    Article Snippet: BHK-21 cells were grown at 37°C in the presence of 5% CO2 .

    Article Title: Application of interferon modulators to overcome partial resistance of human ovarian cancers to VSV-GP oncolytic viral therapy
    Article Snippet: Cell lines BHK21 cells (American Type Culture Collection, Manassas, VA) were maintained in Glasgow minimum essential medium (GMEM) (Gibco, Carlsbad, CA) supplemented with 10% fetal calf serum (FCS; PAA Laboratories, Cölbe, Germany), 5% tryptose phosphate broth (Gibco, Carlsbad, CA), 100 units/ml penicillin (Gibco), and 0.1 mg/ml streptomycin (Gibco).

    Modification:

    Article Title: Mutations in the DI-DII Linker of Human Parainfluenza Virus Type 3 Fusion Protein Result in Diminished Fusion Activity
    Article Snippet: .. Cells and viruses BHK-21 cells, obtained from the American Type Culture Collection, were propagated in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco) supplemented with 1% glutamine, 10% fetal calf serum (FCS) (Gibco), and 1% penicillin-streptomycin (Invitrogen). .. Wild-type (wt) vaccinia virus was used to quantify cell fusion, while recombinant vaccinia virus vTF7-3, a generous gift from Dr. Bernard Moss, was maintained in BHK-21 cells to provide T7 RNA polymerase in the vaccinia-T7 RNA polymerase expression system [ ].

    Article Title: Glycoprotein B Cleavage Is Important for Murid Herpesvirus 4 To Infect Myeloid Cells
    Article Snippet: .. BHK-21, NMuMG, NS0, and RAW 264.7 cells (all from ATCC) were grown in Dulbecco's modified Eagle's medium (DMEM; high glucose; PAA Laboratories) with 2 mM l -glutamine (PAA Laboratories), 100 U/ml penicillin and 100 μg/ml streptomycin (PAA Laboratories), and 10% fetal bovine serum (FBS; PAA Laboratories or Amimed) (complete medium). .. Peritoneal cavity cells were isolated from BALB/c mice by peritoneal lavage and cultivated in RPMI 1640 (PAA Laboratories) with 10% heat-inactivated (56°C, 30 min) FBS (PAA Laboratories or Amimed), 2 mM l -glutamine (PAA Laboratories), and 50 U/ml penicillin and 50 μg/ml streptomycin (PAA Laboratories).

    Article Title: A Novel Dengue Virus Inhibitor, BP13944, Discovered by High-Throughput Screening with Dengue Virus Replicon Cells Selects for Resistance in the Viral NS2B/NS3 Protease
    Article Snippet: .. BHK-21 cells (ATCC CCL-10) were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 4.5 g/liter glucose and 5% fetal bovine serum (FBS). .. The cells were cultured at 37°C in a 5% CO2 incubator.

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  • 85
    ATCC bhk 21 cells
    S.A.AR86, as well as other Sindbis-group alphaviruses, replicates in bone- and joint-associated tissues. (A) Six-week-old female CD-1 mice were infected with 10 3 PFU of s55 i.v. Three days postinfection mice were sacrificed, exsanguinated, and perfused with PBS, and their femurs and quadriceps muscles were removed by dissection and titrated for infectious virus on <t>BHK-21</t> cells. Data are shown as log PFU per gram of tissue for femur and muscle and as PFU per milliliter for serum. Each bar represents a single animal. The arrow indicates the limit of detection, and asterisks denote samples below the limit of detection. Data shown are from one of four experiments. (B) Six-week-old female CD-1 mice were infected i.v. with 10 3 PFU of s51 and sacrificed 3 days postinfection, and right femurs were removed for virus titration. Bone marrow was aspirated from the diaphyses of the femurs using 0.4 ml of PBS–1% donor calf serum per femur. Aspirates were freeze-thawed and titrated for infectious virus by plaque assay. Following marrow aspiration, the remaining femoral tissue was processed as for panel A and titrated for infectious virus by plaque assay. Titer is shown as total PFU per marrow aspirate (Asp) or femur (without aspirated marrow), with each bar representing a single animal. The arrow indicates the limit of detection, and asterisks denote samples with titers below the limit of detection. Shown is one of three comparable experiments. (C) Six-week-old female CD-1 mice were infected i.v. with 10 3 PFU of the virus s55, TR339, or TRSB. Three days postinfection mice were sacrificed and both femurs were removed for virus titration. Femurs were processed and titrated for infectious virus as for panel A. Each bar represents results from a single animal. The arrow indicates the limit of detection. N/D, not done. Shown is one of two comparable experiments.
    Bhk 21 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 85/100, based on 52 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bhk 21 cells/product/ATCC
    Average 85 stars, based on 52 article reviews
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    85
    ATCC treatments bhk 21 cells
    Cytotoxic effect of Fibrillar BSAs . Cell viability was determined by the MTT assay. (A) <t>BHK-21</t> cells were treated for 8 h with various concentrations of G-BSA, BSA-S75, BSA-S200, BSA-Zwit, or BSA-HW55S as indicated in serum-free medium. (B) BHK-21 cells were treated with increasing concentrations of Aβ 25–35 in serum-free medium for 8 h. (C) BHK-21 cells were treated with increasing concentrations of Aβ 25–35 or F-BSA (BSA-S200) in serum-free medium for 24 h. (D) BHK-21 cells were treated with or without increasing concentrations of G-BSA (BSA) for 1 h, then incubated with 0.5 μM F-BSA (BSA-S200) in serum-free medium for 8 h. Data are means ± S.D. (n = 3) of percentage of cell survival as determined by the MTT assays.
    Treatments Bhk 21 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/treatments bhk 21 cells/product/ATCC
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    99
    ATCC viruses bhk 21 cells
    Dye transfer directed by the wt or mutated F and HPIV3 HN proteins. R18-labeled RBCs were bound at 4°C to <t>BHK-21</t> cells coexpressing the wt or mutanted F and HN proteins. Cells were incubated for 60 min at 37°C to allow membrane fusion, images were acquired by using fluorescent microscopy. (A) Representative images of dye transfer. White arrows indicate the lipid mixing events. (B) Quantification of the events of dye transfer. The extent of dye tranfer is expressed as the average number of R18 lipid dye tranfer events of six microscopic fields. The means and standard errors are from six microscopic fields (** P
    Viruses Bhk 21 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    S.A.AR86, as well as other Sindbis-group alphaviruses, replicates in bone- and joint-associated tissues. (A) Six-week-old female CD-1 mice were infected with 10 3 PFU of s55 i.v. Three days postinfection mice were sacrificed, exsanguinated, and perfused with PBS, and their femurs and quadriceps muscles were removed by dissection and titrated for infectious virus on BHK-21 cells. Data are shown as log PFU per gram of tissue for femur and muscle and as PFU per milliliter for serum. Each bar represents a single animal. The arrow indicates the limit of detection, and asterisks denote samples below the limit of detection. Data shown are from one of four experiments. (B) Six-week-old female CD-1 mice were infected i.v. with 10 3 PFU of s51 and sacrificed 3 days postinfection, and right femurs were removed for virus titration. Bone marrow was aspirated from the diaphyses of the femurs using 0.4 ml of PBS–1% donor calf serum per femur. Aspirates were freeze-thawed and titrated for infectious virus by plaque assay. Following marrow aspiration, the remaining femoral tissue was processed as for panel A and titrated for infectious virus by plaque assay. Titer is shown as total PFU per marrow aspirate (Asp) or femur (without aspirated marrow), with each bar representing a single animal. The arrow indicates the limit of detection, and asterisks denote samples with titers below the limit of detection. Shown is one of three comparable experiments. (C) Six-week-old female CD-1 mice were infected i.v. with 10 3 PFU of the virus s55, TR339, or TRSB. Three days postinfection mice were sacrificed and both femurs were removed for virus titration. Femurs were processed and titrated for infectious virus as for panel A. Each bar represents results from a single animal. The arrow indicates the limit of detection. N/D, not done. Shown is one of two comparable experiments.

    Journal: Journal of Virology

    Article Title: Sindbis-Group Alphavirus Replication in Periosteum and Endosteum of Long Bones in Adult Mice

    doi:

    Figure Lengend Snippet: S.A.AR86, as well as other Sindbis-group alphaviruses, replicates in bone- and joint-associated tissues. (A) Six-week-old female CD-1 mice were infected with 10 3 PFU of s55 i.v. Three days postinfection mice were sacrificed, exsanguinated, and perfused with PBS, and their femurs and quadriceps muscles were removed by dissection and titrated for infectious virus on BHK-21 cells. Data are shown as log PFU per gram of tissue for femur and muscle and as PFU per milliliter for serum. Each bar represents a single animal. The arrow indicates the limit of detection, and asterisks denote samples below the limit of detection. Data shown are from one of four experiments. (B) Six-week-old female CD-1 mice were infected i.v. with 10 3 PFU of s51 and sacrificed 3 days postinfection, and right femurs were removed for virus titration. Bone marrow was aspirated from the diaphyses of the femurs using 0.4 ml of PBS–1% donor calf serum per femur. Aspirates were freeze-thawed and titrated for infectious virus by plaque assay. Following marrow aspiration, the remaining femoral tissue was processed as for panel A and titrated for infectious virus by plaque assay. Titer is shown as total PFU per marrow aspirate (Asp) or femur (without aspirated marrow), with each bar representing a single animal. The arrow indicates the limit of detection, and asterisks denote samples with titers below the limit of detection. Shown is one of three comparable experiments. (C) Six-week-old female CD-1 mice were infected i.v. with 10 3 PFU of the virus s55, TR339, or TRSB. Three days postinfection mice were sacrificed and both femurs were removed for virus titration. Femurs were processed and titrated for infectious virus as for panel A. Each bar represents results from a single animal. The arrow indicates the limit of detection. N/D, not done. Shown is one of two comparable experiments.

    Article Snippet: The tissue homogenate was then clarified by centrifugation and assayed for infectious virus by plaque assay on BHK-21 cells (ATCC CRL 8544) as previously described ( ).

    Techniques: Mouse Assay, Infection, Dissection, Titration, Plaque Assay

    Cytotoxic effect of Fibrillar BSAs . Cell viability was determined by the MTT assay. (A) BHK-21 cells were treated for 8 h with various concentrations of G-BSA, BSA-S75, BSA-S200, BSA-Zwit, or BSA-HW55S as indicated in serum-free medium. (B) BHK-21 cells were treated with increasing concentrations of Aβ 25–35 in serum-free medium for 8 h. (C) BHK-21 cells were treated with increasing concentrations of Aβ 25–35 or F-BSA (BSA-S200) in serum-free medium for 24 h. (D) BHK-21 cells were treated with or without increasing concentrations of G-BSA (BSA) for 1 h, then incubated with 0.5 μM F-BSA (BSA-S200) in serum-free medium for 8 h. Data are means ± S.D. (n = 3) of percentage of cell survival as determined by the MTT assays.

    Journal: BMC Biotechnology

    Article Title: Albumin fibrillization induces apoptosis via integrin/FAK/Akt pathway

    doi: 10.1186/1472-6750-9-2

    Figure Lengend Snippet: Cytotoxic effect of Fibrillar BSAs . Cell viability was determined by the MTT assay. (A) BHK-21 cells were treated for 8 h with various concentrations of G-BSA, BSA-S75, BSA-S200, BSA-Zwit, or BSA-HW55S as indicated in serum-free medium. (B) BHK-21 cells were treated with increasing concentrations of Aβ 25–35 in serum-free medium for 8 h. (C) BHK-21 cells were treated with increasing concentrations of Aβ 25–35 or F-BSA (BSA-S200) in serum-free medium for 24 h. (D) BHK-21 cells were treated with or without increasing concentrations of G-BSA (BSA) for 1 h, then incubated with 0.5 μM F-BSA (BSA-S200) in serum-free medium for 8 h. Data are means ± S.D. (n = 3) of percentage of cell survival as determined by the MTT assays.

    Article Snippet: Cell lines and treatments BHK-21 cells (baby hamster kidney; ATCC CRL-1632) and T47D cells (human breast duct carcinoma; ATCC HTB-133) were maintained at 37°C in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, 100 units/ml penicillin, and 100 μg/ml streptomycin.

    Techniques: MTT Assay, Incubation

    Evaluation of the apoptotic effect of fibrillar BSA . (A) BHK-21 cells were incubated with 1 μM G-BSA (BSA) or F-BSA (BSA-S200) for 3 h. The cells were observed under a fluorescence microscope, and their nuclei were stained with DAPI (magnification in all panels, ×400). (B) BHK-21 cells were incubated with 40 μM Aβ 25–35 for 3 h. The cells were observed under a fluorescence microscope, and their nuclei were stained with DAPI (magnification in all panels, ×400). (C). BHK-21 cells were cultured with 0.8 μM G-BSA (BSA) or F-BSA (BSA-S200) for 15 h in serum-free medium, then underwent caspase-3 activity analysis measured by fluorogenic substrate as described under

    Journal: BMC Biotechnology

    Article Title: Albumin fibrillization induces apoptosis via integrin/FAK/Akt pathway

    doi: 10.1186/1472-6750-9-2

    Figure Lengend Snippet: Evaluation of the apoptotic effect of fibrillar BSA . (A) BHK-21 cells were incubated with 1 μM G-BSA (BSA) or F-BSA (BSA-S200) for 3 h. The cells were observed under a fluorescence microscope, and their nuclei were stained with DAPI (magnification in all panels, ×400). (B) BHK-21 cells were incubated with 40 μM Aβ 25–35 for 3 h. The cells were observed under a fluorescence microscope, and their nuclei were stained with DAPI (magnification in all panels, ×400). (C). BHK-21 cells were cultured with 0.8 μM G-BSA (BSA) or F-BSA (BSA-S200) for 15 h in serum-free medium, then underwent caspase-3 activity analysis measured by fluorogenic substrate as described under "Methods". Data are the mean ± SD of three experiments.

    Article Snippet: Cell lines and treatments BHK-21 cells (baby hamster kidney; ATCC CRL-1632) and T47D cells (human breast duct carcinoma; ATCC HTB-133) were maintained at 37°C in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, 100 units/ml penicillin, and 100 μg/ml streptomycin.

    Techniques: Incubation, Fluorescence, Microscopy, Staining, Cell Culture, Activity Assay

    Fibrillar BSA induced cytotoxicity via the integrin/FAK/Akt pathway . (A) BHK-21 cells were treated with 3 μM F-BSA (BSA-S200) in serum-free medium for the indicated time, and cell lysates were analyzed by western blotting with anti-phospho-FAK(Tyr576/577), anti-phospho-FAK(Tyr397), and anti-phospho-Akt (p-Akt) antibodies. (B) BHK-21 cells were pre-treated for 30 min with or without 1 μM goat IgG or 1 μM goat anti-integrin α5β1 antibody as indicated, then treated with 3 μM F-BSA (BSA-S200) in serum-free medium for 15 min. Cell lysates were analyzed by western blotting with anti-phospho-Akt (p-Akt) and anti-phospho-GSK-3β (p-GSK-3β) antibodies. (C) BHK-21 cells were treated with increasing concentrations of G-BSA (BSA) in serum-free medium as indicated, and cell lysates were analyzed by western blotting with anti-phospho-Akt (p-Akt) antibody. (D) BHK-21 cells were treated with or without 1 μM anti-integrin α5β1 antibody in serum-free medium for 30 min, and cell lysates were analyzed by western blotting with anti-phospho-Akt (p-Akt) antibody.

    Journal: BMC Biotechnology

    Article Title: Albumin fibrillization induces apoptosis via integrin/FAK/Akt pathway

    doi: 10.1186/1472-6750-9-2

    Figure Lengend Snippet: Fibrillar BSA induced cytotoxicity via the integrin/FAK/Akt pathway . (A) BHK-21 cells were treated with 3 μM F-BSA (BSA-S200) in serum-free medium for the indicated time, and cell lysates were analyzed by western blotting with anti-phospho-FAK(Tyr576/577), anti-phospho-FAK(Tyr397), and anti-phospho-Akt (p-Akt) antibodies. (B) BHK-21 cells were pre-treated for 30 min with or without 1 μM goat IgG or 1 μM goat anti-integrin α5β1 antibody as indicated, then treated with 3 μM F-BSA (BSA-S200) in serum-free medium for 15 min. Cell lysates were analyzed by western blotting with anti-phospho-Akt (p-Akt) and anti-phospho-GSK-3β (p-GSK-3β) antibodies. (C) BHK-21 cells were treated with increasing concentrations of G-BSA (BSA) in serum-free medium as indicated, and cell lysates were analyzed by western blotting with anti-phospho-Akt (p-Akt) antibody. (D) BHK-21 cells were treated with or without 1 μM anti-integrin α5β1 antibody in serum-free medium for 30 min, and cell lysates were analyzed by western blotting with anti-phospho-Akt (p-Akt) antibody.

    Article Snippet: Cell lines and treatments BHK-21 cells (baby hamster kidney; ATCC CRL-1632) and T47D cells (human breast duct carcinoma; ATCC HTB-133) were maintained at 37°C in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, 100 units/ml penicillin, and 100 μg/ml streptomycin.

    Techniques: Western Blot

    Dye transfer directed by the wt or mutated F and HPIV3 HN proteins. R18-labeled RBCs were bound at 4°C to BHK-21 cells coexpressing the wt or mutanted F and HN proteins. Cells were incubated for 60 min at 37°C to allow membrane fusion, images were acquired by using fluorescent microscopy. (A) Representative images of dye transfer. White arrows indicate the lipid mixing events. (B) Quantification of the events of dye transfer. The extent of dye tranfer is expressed as the average number of R18 lipid dye tranfer events of six microscopic fields. The means and standard errors are from six microscopic fields (** P

    Journal: PLoS ONE

    Article Title: Mutations in the DI-DII Linker of Human Parainfluenza Virus Type 3 Fusion Protein Result in Diminished Fusion Activity

    doi: 10.1371/journal.pone.0136474

    Figure Lengend Snippet: Dye transfer directed by the wt or mutated F and HPIV3 HN proteins. R18-labeled RBCs were bound at 4°C to BHK-21 cells coexpressing the wt or mutanted F and HN proteins. Cells were incubated for 60 min at 37°C to allow membrane fusion, images were acquired by using fluorescent microscopy. (A) Representative images of dye transfer. White arrows indicate the lipid mixing events. (B) Quantification of the events of dye transfer. The extent of dye tranfer is expressed as the average number of R18 lipid dye tranfer events of six microscopic fields. The means and standard errors are from six microscopic fields (** P

    Article Snippet: Cells and viruses BHK-21 cells, obtained from the American Type Culture Collection, were propagated in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco) supplemented with 1% glutamine, 10% fetal calf serum (FCS) (Gibco), and 1% penicillin-streptomycin (Invitrogen).

    Techniques: Labeling, Incubation, Microscopy

    Syncytium formation in monolayers coexpressing wt or mutated F and HN proteins. After 36 h posttransfection, BHK-21 monolayers transfected with vector alone, wt F alone, wt F and wt HN, or mutant F and wt HN were fixed with methanol and stained with Giemsa stain. (A) Photomicrographs from a representative expreriment. Red arrows indicate syncytia. (B) Quantification of syncytia produced by the mutated F proteins. Values were indicated as percentages of syncytium formation detected in cells transfected with wt F and wt HN proteins and are represented as the mean ± standard deviation (SD) from three separate experiments (* P

    Journal: PLoS ONE

    Article Title: Mutations in the DI-DII Linker of Human Parainfluenza Virus Type 3 Fusion Protein Result in Diminished Fusion Activity

    doi: 10.1371/journal.pone.0136474

    Figure Lengend Snippet: Syncytium formation in monolayers coexpressing wt or mutated F and HN proteins. After 36 h posttransfection, BHK-21 monolayers transfected with vector alone, wt F alone, wt F and wt HN, or mutant F and wt HN were fixed with methanol and stained with Giemsa stain. (A) Photomicrographs from a representative expreriment. Red arrows indicate syncytia. (B) Quantification of syncytia produced by the mutated F proteins. Values were indicated as percentages of syncytium formation detected in cells transfected with wt F and wt HN proteins and are represented as the mean ± standard deviation (SD) from three separate experiments (* P

    Article Snippet: Cells and viruses BHK-21 cells, obtained from the American Type Culture Collection, were propagated in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco) supplemented with 1% glutamine, 10% fetal calf serum (FCS) (Gibco), and 1% penicillin-streptomycin (Invitrogen).

    Techniques: Transfection, Plasmid Preparation, Mutagenesis, Staining, Giemsa Stain, Produced, Standard Deviation

    Growth properties of gB FCS − viruses. (A) Low-MOI growth curve. BHK-21 cells were infected with eGFP + WT virus and ΔFCSv1, ΔFCSv2, and ΔFCSv3 mutant and revertant viruses at an MOI of 0.05 eGFP units/cell. At the indicated times p.i., the cells were lysed by 1 cycle of freeze-thawing and titrated on BHK-21 cells by counting eGFP + cells by flow cytometry. Each point shows the mean ± standard error of the mean from 3 wells. The dashed line shows the sensitivity threshold of virus titration. The titers of the gB FCS − viruses (ΔFCSv1, ΔFCSv2, and ΔFCSv3) were significantly lower than those of the gB FCS + viruses (WT, ΔFCSv1 R, ΔFCSv2 R, and ΔFCSv3 R) on days 3 ( P

    Journal: Journal of Virology

    Article Title: Glycoprotein B Cleavage Is Important for Murid Herpesvirus 4 To Infect Myeloid Cells

    doi: 10.1128/JVI.00709-13

    Figure Lengend Snippet: Growth properties of gB FCS − viruses. (A) Low-MOI growth curve. BHK-21 cells were infected with eGFP + WT virus and ΔFCSv1, ΔFCSv2, and ΔFCSv3 mutant and revertant viruses at an MOI of 0.05 eGFP units/cell. At the indicated times p.i., the cells were lysed by 1 cycle of freeze-thawing and titrated on BHK-21 cells by counting eGFP + cells by flow cytometry. Each point shows the mean ± standard error of the mean from 3 wells. The dashed line shows the sensitivity threshold of virus titration. The titers of the gB FCS − viruses (ΔFCSv1, ΔFCSv2, and ΔFCSv3) were significantly lower than those of the gB FCS + viruses (WT, ΔFCSv1 R, ΔFCSv2 R, and ΔFCSv3 R) on days 3 ( P

    Article Snippet: BHK-21, NMuMG, NS0, and RAW 264.7 cells (all from ATCC) were grown in Dulbecco's modified Eagle's medium (DMEM; high glucose; PAA Laboratories) with 2 mM l -glutamine (PAA Laboratories), 100 U/ml penicillin and 100 μg/ml streptomycin (PAA Laboratories), and 10% fetal bovine serum (FBS; PAA Laboratories or Amimed) (complete medium).

    Techniques: Infection, Mutagenesis, Flow Cytometry, Cytometry, Titration

    Susceptibility of gB FCS − viruses to gB-directed neutralization. eGFP + WT, ΔFCSv1 and ΔFCSv3 mutant, and ΔFCSv1 revertant viruses (MOI of 0.5 eGFP units/cell) were incubated (2 h, RT) with the gB-specific neutralizing MAbs SC-9E8 and SC-9A5 blocking viral membrane fusion or the gH/gL-specific MAb 8F10 blocking cell binding. The viruses were then added to BHK-21 cells and incubated for 18 h at 37°C in the presence of 100 μg/ml PAA. The eGFP + cells were enumerated by flow cytometry and are shown relative to untreated virus. Each point shows the mean ± standard error of the mean from 3 wells. The gB FCS − viruses (ΔFCSv1 and ΔFCSv3) were neutralized significantly better than the gB FCS + viruses (WT and ΔFCSv1 R) with MAb SC-9E8 at antibody doses of 16.7 ( P

    Journal: Journal of Virology

    Article Title: Glycoprotein B Cleavage Is Important for Murid Herpesvirus 4 To Infect Myeloid Cells

    doi: 10.1128/JVI.00709-13

    Figure Lengend Snippet: Susceptibility of gB FCS − viruses to gB-directed neutralization. eGFP + WT, ΔFCSv1 and ΔFCSv3 mutant, and ΔFCSv1 revertant viruses (MOI of 0.5 eGFP units/cell) were incubated (2 h, RT) with the gB-specific neutralizing MAbs SC-9E8 and SC-9A5 blocking viral membrane fusion or the gH/gL-specific MAb 8F10 blocking cell binding. The viruses were then added to BHK-21 cells and incubated for 18 h at 37°C in the presence of 100 μg/ml PAA. The eGFP + cells were enumerated by flow cytometry and are shown relative to untreated virus. Each point shows the mean ± standard error of the mean from 3 wells. The gB FCS − viruses (ΔFCSv1 and ΔFCSv3) were neutralized significantly better than the gB FCS + viruses (WT and ΔFCSv1 R) with MAb SC-9E8 at antibody doses of 16.7 ( P

    Article Snippet: BHK-21, NMuMG, NS0, and RAW 264.7 cells (all from ATCC) were grown in Dulbecco's modified Eagle's medium (DMEM; high glucose; PAA Laboratories) with 2 mM l -glutamine (PAA Laboratories), 100 U/ml penicillin and 100 μg/ml streptomycin (PAA Laboratories), and 10% fetal bovine serum (FBS; PAA Laboratories or Amimed) (complete medium).

    Techniques: Neutralization, Mutagenesis, Incubation, Blocking Assay, Binding Assay, Flow Cytometry, Cytometry

    Antigenic conformation of uncleaved gB. (A) BHK-21 cells were infected with eGFP + WT virus and ΔFCSv1, ΔFCSv2, and ΔFCSv3 mutant and revertant viruses at an MOI of 2 PFU/cell. Eighteen hours later, cells were stained with MAbs BN-1A7 and SC-9E8 recognizing prefusion gB, MAb MG-1A12 recognizing postfusion gB, MAb T2C12 recognizing gH/gL, and MAb BN-3A4 recognizing gp150. Bound antibody was detected with an Alexa Fluor 633-conjugated secondary antibody and flow cytometry (open histograms). The gray histograms show staining with secondary antibody only. Detection of gH/gL and gp150 served as the control for equivalent infection of cells. Equivalent data were obtained in a repeat experiment. (B) NMuMG cells were incubated with eGFP + WT, ΔFCSv1 and ΔFCSv3 mutant, and ΔFCSv1 revertant viruses at an MOI of 15 eGFP units/cell (2 h, 4°C). After 3 washes in ice-cold PBS to remove unbound virions, the cells were fixed either directly or after further incubations at 37°C to allow virion endocytosis. The cells were then incubated with MAb BN-1A7 recognizing prefusion gB and MAb MG-1A12 recognizing postfusion gB. Bound antibody was detected with an alkaline phosphatase-conjugated secondary antibody, incubation with p -nitrophenyl phosphate substrate, and measurement of the absorbance at 405 nm ( A 405 ). The bars show means ± standard errors of the means from 6 wells. Equivalent data were obtained in 3 further experiments.

    Journal: Journal of Virology

    Article Title: Glycoprotein B Cleavage Is Important for Murid Herpesvirus 4 To Infect Myeloid Cells

    doi: 10.1128/JVI.00709-13

    Figure Lengend Snippet: Antigenic conformation of uncleaved gB. (A) BHK-21 cells were infected with eGFP + WT virus and ΔFCSv1, ΔFCSv2, and ΔFCSv3 mutant and revertant viruses at an MOI of 2 PFU/cell. Eighteen hours later, cells were stained with MAbs BN-1A7 and SC-9E8 recognizing prefusion gB, MAb MG-1A12 recognizing postfusion gB, MAb T2C12 recognizing gH/gL, and MAb BN-3A4 recognizing gp150. Bound antibody was detected with an Alexa Fluor 633-conjugated secondary antibody and flow cytometry (open histograms). The gray histograms show staining with secondary antibody only. Detection of gH/gL and gp150 served as the control for equivalent infection of cells. Equivalent data were obtained in a repeat experiment. (B) NMuMG cells were incubated with eGFP + WT, ΔFCSv1 and ΔFCSv3 mutant, and ΔFCSv1 revertant viruses at an MOI of 15 eGFP units/cell (2 h, 4°C). After 3 washes in ice-cold PBS to remove unbound virions, the cells were fixed either directly or after further incubations at 37°C to allow virion endocytosis. The cells were then incubated with MAb BN-1A7 recognizing prefusion gB and MAb MG-1A12 recognizing postfusion gB. Bound antibody was detected with an alkaline phosphatase-conjugated secondary antibody, incubation with p -nitrophenyl phosphate substrate, and measurement of the absorbance at 405 nm ( A 405 ). The bars show means ± standard errors of the means from 6 wells. Equivalent data were obtained in 3 further experiments.

    Article Snippet: BHK-21, NMuMG, NS0, and RAW 264.7 cells (all from ATCC) were grown in Dulbecco's modified Eagle's medium (DMEM; high glucose; PAA Laboratories) with 2 mM l -glutamine (PAA Laboratories), 100 U/ml penicillin and 100 μg/ml streptomycin (PAA Laboratories), and 10% fetal bovine serum (FBS; PAA Laboratories or Amimed) (complete medium).

    Techniques: Infection, Mutagenesis, Staining, Flow Cytometry, Cytometry, Incubation

    Kinetics of virus entry in fibroblasts, epithelial cells, and macrophages. (A) BHK-21 fibroblasts were exposed at 37°C to eGFP + WT, ΔFCSv1 and ΔFCSv3 mutant, and ΔFCSv1 revertant viruses at an MOI of 1 eGFP units/cell in the presence of 100 μg/ml PAA for the times indicated on the x axis. The cells were then washed with PBS to remove unbound virions or with phosphate-citrate buffer (pH 3) (acid wash) to inactivate all extracellular virions, followed by incubation at 37°C in the presence of 100 μg/ml PAA until 24 h p.i. The proportion of eGFP + cells was then determined by flow cytometry. Each point shows the mean ± standard error of the mean from 3 wells. The incubation times required for half-maximal infection levels ( t 50% ) were as follows (mean ± standard error of the mean): PBS wash, 2 ± 0.05 h for gB FCS + (WT and ΔFCSv1 R) and 2.3 ± 0.1 h for gB FCS − (ΔFCSv1 and ΔFCSv3) viruses; acid wash, 4.1 ± 0.08 h for gB FCS + and 4.7 ± 0.15 h for gB FCS − viruses. For acid wash, the difference in t 50% values was statistically significant ( P

    Journal: Journal of Virology

    Article Title: Glycoprotein B Cleavage Is Important for Murid Herpesvirus 4 To Infect Myeloid Cells

    doi: 10.1128/JVI.00709-13

    Figure Lengend Snippet: Kinetics of virus entry in fibroblasts, epithelial cells, and macrophages. (A) BHK-21 fibroblasts were exposed at 37°C to eGFP + WT, ΔFCSv1 and ΔFCSv3 mutant, and ΔFCSv1 revertant viruses at an MOI of 1 eGFP units/cell in the presence of 100 μg/ml PAA for the times indicated on the x axis. The cells were then washed with PBS to remove unbound virions or with phosphate-citrate buffer (pH 3) (acid wash) to inactivate all extracellular virions, followed by incubation at 37°C in the presence of 100 μg/ml PAA until 24 h p.i. The proportion of eGFP + cells was then determined by flow cytometry. Each point shows the mean ± standard error of the mean from 3 wells. The incubation times required for half-maximal infection levels ( t 50% ) were as follows (mean ± standard error of the mean): PBS wash, 2 ± 0.05 h for gB FCS + (WT and ΔFCSv1 R) and 2.3 ± 0.1 h for gB FCS − (ΔFCSv1 and ΔFCSv3) viruses; acid wash, 4.1 ± 0.08 h for gB FCS + and 4.7 ± 0.15 h for gB FCS − viruses. For acid wash, the difference in t 50% values was statistically significant ( P

    Article Snippet: BHK-21, NMuMG, NS0, and RAW 264.7 cells (all from ATCC) were grown in Dulbecco's modified Eagle's medium (DMEM; high glucose; PAA Laboratories) with 2 mM l -glutamine (PAA Laboratories), 100 U/ml penicillin and 100 μg/ml streptomycin (PAA Laboratories), and 10% fetal bovine serum (FBS; PAA Laboratories or Amimed) (complete medium).

    Techniques: Mutagenesis, Incubation, Flow Cytometry, Cytometry, Infection

    Comparative analysis of virus entry in fibroblasts, epithelial cells, myeloma cells, macrophages, and DCs. (A and B) BHK-21 fibroblasts (A) or NMuMG epithelial cells (B) were infected with eGFP + WT, ΔFCSv1 and ΔFCSv3 mutant, and ΔFCSv1 revertant viruses at an MOI of 0.5 eGFP units/cell. After incubation in the presence of 100 μg/ml PAA for 18 h at 37°C, the proportion of eGFP + cells was determined by flow cytometry. The bars show the means ± standard errors of the means from 3 wells. Equivalent data were obtained in 3 further experiments. (C) NS0 myeloma cells were infected as for panel A but at an MOI of 100 eGFP units/cell. The bars show the means ± standard errors of the means from 3 wells. The values for the gB FCS − viruses (ΔFCSv1 and ΔFCSv3) were significantly lower than those for the gB FCS + viruses (WT and ΔFCSv1 R) ( P

    Journal: Journal of Virology

    Article Title: Glycoprotein B Cleavage Is Important for Murid Herpesvirus 4 To Infect Myeloid Cells

    doi: 10.1128/JVI.00709-13

    Figure Lengend Snippet: Comparative analysis of virus entry in fibroblasts, epithelial cells, myeloma cells, macrophages, and DCs. (A and B) BHK-21 fibroblasts (A) or NMuMG epithelial cells (B) were infected with eGFP + WT, ΔFCSv1 and ΔFCSv3 mutant, and ΔFCSv1 revertant viruses at an MOI of 0.5 eGFP units/cell. After incubation in the presence of 100 μg/ml PAA for 18 h at 37°C, the proportion of eGFP + cells was determined by flow cytometry. The bars show the means ± standard errors of the means from 3 wells. Equivalent data were obtained in 3 further experiments. (C) NS0 myeloma cells were infected as for panel A but at an MOI of 100 eGFP units/cell. The bars show the means ± standard errors of the means from 3 wells. The values for the gB FCS − viruses (ΔFCSv1 and ΔFCSv3) were significantly lower than those for the gB FCS + viruses (WT and ΔFCSv1 R) ( P

    Article Snippet: BHK-21, NMuMG, NS0, and RAW 264.7 cells (all from ATCC) were grown in Dulbecco's modified Eagle's medium (DMEM; high glucose; PAA Laboratories) with 2 mM l -glutamine (PAA Laboratories), 100 U/ml penicillin and 100 μg/ml streptomycin (PAA Laboratories), and 10% fetal bovine serum (FBS; PAA Laboratories or Amimed) (complete medium).

    Techniques: Infection, Mutagenesis, Incubation, Flow Cytometry, Cytometry