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  • 89
    Stratagene pcmv tag2a
    Mybbp1a was identified as a novel candidate binding protein for TBP-2. ( a ) Purification scheme for the TBP-2 containing protein complex in β-cell nuclear extracts. TBP-2 binding proteins were purified from INS-1 nuclear extracts by using TBP-2 protein-immobilized (TBP-2) or control beads. ( b ) Identification of the TBP-2 complex. Silver staining was performed. Numbers indicate individual candidate proteins interacting with TBP-2. ( c ) Eluted proteins were analysed by immuno blotting (IB) for Mybbp1a and TBP-2 antibody. ( d ) Co-immunoprecipitation analyses. Input (left, 5% lysate) and anti-Myc immunoprecipitates (right, IP:Myc) from HEK293 cells transfected with <t>pCMV-tag2A</t> and pCMV-tag3B vector (−) or FLAG-HA-Mybbp1a and Myc-TBP-2 vector (+) were analysed by immunoblotting (IB) with antibodies to FLAG, Myc and β-actin. The positions for Mybbp1a (p160; p160MBP) and Mybbp1a (p140; p140MBP) (upper), TBP-2 (middle) and β-actin (lower) are shown. ( e ) Luciferase activity of the UCP-2 -86 enhancer region. INS-1 cells were transfected with PGL4.10 UCP-2 -86 or PGL4.10-luciferase reporter plasmid and each expression plasmid for PGC-1α, Mybbp1a and TBP-2, as indicated. Luciferase reporter activity was normalized by Renilla luciferase activity. ( f ) A schematic model of TBP-2 function in β-cells. Mybbp1a binds PGC-1α (inactive form) and inhibits UCP-2 transcriptional activity. Induced TBP-2 interacts with Mybbp1a and releases PGC-1α from Mybbp1a, facilitating PGC-1α recruitment on the UCP-2 promoter region. Data are presented as mean±s.d. * P
    Pcmv Tag2a, supplied by Stratagene, used in various techniques. Bioz Stars score: 89/100, based on 239 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies pcmv tag2a
    Mybbp1a was identified as a novel candidate binding protein for TBP-2. ( a ) Purification scheme for the TBP-2 containing protein complex in β-cell nuclear extracts. TBP-2 binding proteins were purified from INS-1 nuclear extracts by using TBP-2 protein-immobilized (TBP-2) or control beads. ( b ) Identification of the TBP-2 complex. Silver staining was performed. Numbers indicate individual candidate proteins interacting with TBP-2. ( c ) Eluted proteins were analysed by immuno blotting (IB) for Mybbp1a and TBP-2 antibody. ( d ) Co-immunoprecipitation analyses. Input (left, 5% lysate) and anti-Myc immunoprecipitates (right, IP:Myc) from HEK293 cells transfected with <t>pCMV-tag2A</t> and pCMV-tag3B vector (−) or FLAG-HA-Mybbp1a and Myc-TBP-2 vector (+) were analysed by immunoblotting (IB) with antibodies to FLAG, Myc and β-actin. The positions for Mybbp1a (p160; p160MBP) and Mybbp1a (p140; p140MBP) (upper), TBP-2 (middle) and β-actin (lower) are shown. ( e ) Luciferase activity of the UCP-2 -86 enhancer region. INS-1 cells were transfected with PGL4.10 UCP-2 -86 or PGL4.10-luciferase reporter plasmid and each expression plasmid for PGC-1α, Mybbp1a and TBP-2, as indicated. Luciferase reporter activity was normalized by Renilla luciferase activity. ( f ) A schematic model of TBP-2 function in β-cells. Mybbp1a binds PGC-1α (inactive form) and inhibits UCP-2 transcriptional activity. Induced TBP-2 interacts with Mybbp1a and releases PGC-1α from Mybbp1a, facilitating PGC-1α recruitment on the UCP-2 promoter region. Data are presented as mean±s.d. * P
    Pcmv Tag2a, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 90/100, based on 134 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Stratagene pcmv tag2a vector
    Ectopic expression of TBP-2 in the late stage of HTLV-I-transformed T cells inhibits cell growth and causes apoptosis. I-ED40515T cells ( a ) and I-ATL43T cells ( b ) were transfected with 2 μg <t>pCMV-tag2A-TBP2</t> or empty vector pCMV-tag2A, respectively. After 48 h, immunoblotting was performed using an anti-Flag antibody to detect Flag-tagged TBP-2 expression (upper panel) and other corresponding antibodies to detect cleaved caspase-3, PARP, β-actin, respectively (middle panel). Flow cytometry assay was performed to detect early apoptotic proportion (lower panel). Data are shown as mean±s.d. ( n =3). *** P
    Pcmv Tag2a Vector, supplied by Stratagene, used in various techniques. Bioz Stars score: 88/100, based on 89 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Stratagene flag tagged pcmv tag2a
    Ectopic expression of TBP-2 in the late stage of HTLV-I-transformed T cells inhibits cell growth and causes apoptosis. I-ED40515T cells ( a ) and I-ATL43T cells ( b ) were transfected with 2 μg <t>pCMV-tag2A-TBP2</t> or empty vector pCMV-tag2A, respectively. After 48 h, immunoblotting was performed using an anti-Flag antibody to detect Flag-tagged TBP-2 expression (upper panel) and other corresponding antibodies to detect cleaved caspase-3, PARP, β-actin, respectively (middle panel). Flow cytometry assay was performed to detect early apoptotic proportion (lower panel). Data are shown as mean±s.d. ( n =3). *** P
    Flag Tagged Pcmv Tag2a, supplied by Stratagene, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Agilent technologies pcmv tag2a vector
    Ectopic expression of TBP-2 in the late stage of HTLV-I-transformed T cells inhibits cell growth and causes apoptosis. I-ED40515T cells ( a ) and I-ATL43T cells ( b ) were transfected with 2 μg <t>pCMV-tag2A-TBP2</t> or empty vector pCMV-tag2A, respectively. After 48 h, immunoblotting was performed using an anti-Flag antibody to detect Flag-tagged TBP-2 expression (upper panel) and other corresponding antibodies to detect cleaved caspase-3, PARP, β-actin, respectively (middle panel). Flow cytometry assay was performed to detect early apoptotic proportion (lower panel). Data are shown as mean±s.d. ( n =3). *** P
    Pcmv Tag2a Vector, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 92/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Thermo Fisher pcmv tag2a
    Ectopic expression of TBP-2 in the late stage of HTLV-I-transformed T cells inhibits cell growth and causes apoptosis. I-ED40515T cells ( a ) and I-ATL43T cells ( b ) were transfected with 2 μg <t>pCMV-tag2A-TBP2</t> or empty vector pCMV-tag2A, respectively. After 48 h, immunoblotting was performed using an anti-Flag antibody to detect Flag-tagged TBP-2 expression (upper panel) and other corresponding antibodies to detect cleaved caspase-3, PARP, β-actin, respectively (middle panel). Flow cytometry assay was performed to detect early apoptotic proportion (lower panel). Data are shown as mean±s.d. ( n =3). *** P
    Pcmv Tag2a, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene ecorv xhoi cut pcmv tag2a
    Ectopic expression of TBP-2 in the late stage of HTLV-I-transformed T cells inhibits cell growth and causes apoptosis. I-ED40515T cells ( a ) and I-ATL43T cells ( b ) were transfected with 2 μg <t>pCMV-tag2A-TBP2</t> or empty vector pCMV-tag2A, respectively. After 48 h, immunoblotting was performed using an anti-Flag antibody to detect Flag-tagged TBP-2 expression (upper panel) and other corresponding antibodies to detect cleaved caspase-3, PARP, β-actin, respectively (middle panel). Flow cytometry assay was performed to detect early apoptotic proportion (lower panel). Data are shown as mean±s.d. ( n =3). *** P
    Ecorv Xhoi Cut Pcmv Tag2a, supplied by Stratagene, used in various techniques. Bioz Stars score: 85/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Agilent technologies pcmv tag2a jph3 plasmid
    Ectopic expression of TBP-2 in the late stage of HTLV-I-transformed T cells inhibits cell growth and causes apoptosis. I-ED40515T cells ( a ) and I-ATL43T cells ( b ) were transfected with 2 μg <t>pCMV-tag2A-TBP2</t> or empty vector pCMV-tag2A, respectively. After 48 h, immunoblotting was performed using an anti-Flag antibody to detect Flag-tagged TBP-2 expression (upper panel) and other corresponding antibodies to detect cleaved caspase-3, PARP, β-actin, respectively (middle panel). Flow cytometry assay was performed to detect early apoptotic proportion (lower panel). Data are shown as mean±s.d. ( n =3). *** P
    Pcmv Tag2a Jph3 Plasmid, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 91/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene mammalian pcmv tag2a
    Ectopic expression of TBP-2 in the late stage of HTLV-I-transformed T cells inhibits cell growth and causes apoptosis. I-ED40515T cells ( a ) and I-ATL43T cells ( b ) were transfected with 2 μg <t>pCMV-tag2A-TBP2</t> or empty vector pCMV-tag2A, respectively. After 48 h, immunoblotting was performed using an anti-Flag antibody to detect Flag-tagged TBP-2 expression (upper panel) and other corresponding antibodies to detect cleaved caspase-3, PARP, β-actin, respectively (middle panel). Flow cytometry assay was performed to detect early apoptotic proportion (lower panel). Data are shown as mean±s.d. ( n =3). *** P
    Mammalian Pcmv Tag2a, supplied by Stratagene, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies pcmv tag2a mgfp
    Ectopic expression of TBP-2 in the late stage of HTLV-I-transformed T cells inhibits cell growth and causes apoptosis. I-ED40515T cells ( a ) and I-ATL43T cells ( b ) were transfected with 2 μg <t>pCMV-tag2A-TBP2</t> or empty vector pCMV-tag2A, respectively. After 48 h, immunoblotting was performed using an anti-Flag antibody to detect Flag-tagged TBP-2 expression (upper panel) and other corresponding antibodies to detect cleaved caspase-3, PARP, β-actin, respectively (middle panel). Flow cytometry assay was performed to detect early apoptotic proportion (lower panel). Data are shown as mean±s.d. ( n =3). *** P
    Pcmv Tag2a Mgfp, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 90/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Inserm Transfert pcmv tag2a cux1
    Ectopic expression of TBP-2 in the late stage of HTLV-I-transformed T cells inhibits cell growth and causes apoptosis. I-ED40515T cells ( a ) and I-ATL43T cells ( b ) were transfected with 2 μg <t>pCMV-tag2A-TBP2</t> or empty vector pCMV-tag2A, respectively. After 48 h, immunoblotting was performed using an anti-Flag antibody to detect Flag-tagged TBP-2 expression (upper panel) and other corresponding antibodies to detect cleaved caspase-3, PARP, β-actin, respectively (middle panel). Flow cytometry assay was performed to detect early apoptotic proportion (lower panel). Data are shown as mean±s.d. ( n =3). *** P
    Pcmv Tag2a Cux1, supplied by Inserm Transfert, used in various techniques. Bioz Stars score: 91/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Stratagene mammalian expression vector pcmv tag2a
    Ectopic expression of TBP-2 in the late stage of HTLV-I-transformed T cells inhibits cell growth and causes apoptosis. I-ED40515T cells ( a ) and I-ATL43T cells ( b ) were transfected with 2 μg <t>pCMV-tag2A-TBP2</t> or empty vector pCMV-tag2A, respectively. After 48 h, immunoblotting was performed using an anti-Flag antibody to detect Flag-tagged TBP-2 expression (upper panel) and other corresponding antibodies to detect cleaved caspase-3, PARP, β-actin, respectively (middle panel). Flow cytometry assay was performed to detect early apoptotic proportion (lower panel). Data are shown as mean±s.d. ( n =3). *** P
    Mammalian Expression Vector Pcmv Tag2a, supplied by Stratagene, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology control plasmid pcmv tag2a
    Ectopic expression of TBP-2 in the late stage of HTLV-I-transformed T cells inhibits cell growth and causes apoptosis. I-ED40515T cells ( a ) and I-ATL43T cells ( b ) were transfected with 2 μg <t>pCMV-tag2A-TBP2</t> or empty vector pCMV-tag2A, respectively. After 48 h, immunoblotting was performed using an anti-Flag antibody to detect Flag-tagged TBP-2 expression (upper panel) and other corresponding antibodies to detect cleaved caspase-3, PARP, β-actin, respectively (middle panel). Flow cytometry assay was performed to detect early apoptotic proportion (lower panel). Data are shown as mean±s.d. ( n =3). *** P
    Control Plasmid Pcmv Tag2a, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene mammalian expression vector pcmv tag2a flag vector
    Ectopic expression of TBP-2 in the late stage of HTLV-I-transformed T cells inhibits cell growth and causes apoptosis. I-ED40515T cells ( a ) and I-ATL43T cells ( b ) were transfected with 2 μg <t>pCMV-tag2A-TBP2</t> or empty vector pCMV-tag2A, respectively. After 48 h, immunoblotting was performed using an anti-Flag antibody to detect Flag-tagged TBP-2 expression (upper panel) and other corresponding antibodies to detect cleaved caspase-3, PARP, β-actin, respectively (middle panel). Flow cytometry assay was performed to detect early apoptotic proportion (lower panel). Data are shown as mean±s.d. ( n =3). *** P
    Mammalian Expression Vector Pcmv Tag2a Flag Vector, supplied by Stratagene, used in various techniques. Bioz Stars score: 85/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene hindiii sali cut pcmv tag2a vector
    Ectopic expression of TBP-2 in the late stage of HTLV-I-transformed T cells inhibits cell growth and causes apoptosis. I-ED40515T cells ( a ) and I-ATL43T cells ( b ) were transfected with 2 μg <t>pCMV-tag2A-TBP2</t> or empty vector pCMV-tag2A, respectively. After 48 h, immunoblotting was performed using an anti-Flag antibody to detect Flag-tagged TBP-2 expression (upper panel) and other corresponding antibodies to detect cleaved caspase-3, PARP, β-actin, respectively (middle panel). Flow cytometry assay was performed to detect early apoptotic proportion (lower panel). Data are shown as mean±s.d. ( n =3). *** P
    Hindiii Sali Cut Pcmv Tag2a Vector, supplied by Stratagene, used in various techniques. Bioz Stars score: 85/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Thermo Fisher mammalian expression vector pcmv tag2a
    Ectopic expression of TBP-2 in the late stage of HTLV-I-transformed T cells inhibits cell growth and causes apoptosis. I-ED40515T cells ( a ) and I-ATL43T cells ( b ) were transfected with 2 μg <t>pCMV-tag2A-TBP2</t> or empty vector pCMV-tag2A, respectively. After 48 h, immunoblotting was performed using an anti-Flag antibody to detect Flag-tagged TBP-2 expression (upper panel) and other corresponding antibodies to detect cleaved caspase-3, PARP, β-actin, respectively (middle panel). Flow cytometry assay was performed to detect early apoptotic proportion (lower panel). Data are shown as mean±s.d. ( n =3). *** P
    Mammalian Expression Vector Pcmv Tag2a, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Millipore pcmv tag 2a
    Ectopic expression of TBP-2 in the late stage of HTLV-I-transformed T cells inhibits cell growth and causes apoptosis. I-ED40515T cells ( a ) and I-ATL43T cells ( b ) were transfected with 2 μg <t>pCMV-tag2A-TBP2</t> or empty vector pCMV-tag2A, respectively. After 48 h, immunoblotting was performed using an anti-Flag antibody to detect Flag-tagged TBP-2 expression (upper panel) and other corresponding antibodies to detect cleaved caspase-3, PARP, β-actin, respectively (middle panel). Flow cytometry assay was performed to detect early apoptotic proportion (lower panel). Data are shown as mean±s.d. ( n =3). *** P
    Pcmv Tag 2a, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Mybbp1a was identified as a novel candidate binding protein for TBP-2. ( a ) Purification scheme for the TBP-2 containing protein complex in β-cell nuclear extracts. TBP-2 binding proteins were purified from INS-1 nuclear extracts by using TBP-2 protein-immobilized (TBP-2) or control beads. ( b ) Identification of the TBP-2 complex. Silver staining was performed. Numbers indicate individual candidate proteins interacting with TBP-2. ( c ) Eluted proteins were analysed by immuno blotting (IB) for Mybbp1a and TBP-2 antibody. ( d ) Co-immunoprecipitation analyses. Input (left, 5% lysate) and anti-Myc immunoprecipitates (right, IP:Myc) from HEK293 cells transfected with pCMV-tag2A and pCMV-tag3B vector (−) or FLAG-HA-Mybbp1a and Myc-TBP-2 vector (+) were analysed by immunoblotting (IB) with antibodies to FLAG, Myc and β-actin. The positions for Mybbp1a (p160; p160MBP) and Mybbp1a (p140; p140MBP) (upper), TBP-2 (middle) and β-actin (lower) are shown. ( e ) Luciferase activity of the UCP-2 -86 enhancer region. INS-1 cells were transfected with PGL4.10 UCP-2 -86 or PGL4.10-luciferase reporter plasmid and each expression plasmid for PGC-1α, Mybbp1a and TBP-2, as indicated. Luciferase reporter activity was normalized by Renilla luciferase activity. ( f ) A schematic model of TBP-2 function in β-cells. Mybbp1a binds PGC-1α (inactive form) and inhibits UCP-2 transcriptional activity. Induced TBP-2 interacts with Mybbp1a and releases PGC-1α from Mybbp1a, facilitating PGC-1α recruitment on the UCP-2 promoter region. Data are presented as mean±s.d. * P

    Journal: Nature Communications

    Article Title: Disruption of TBP-2 ameliorates insulin sensitivity and secretion without affecting obesity

    doi: 10.1038/ncomms1127

    Figure Lengend Snippet: Mybbp1a was identified as a novel candidate binding protein for TBP-2. ( a ) Purification scheme for the TBP-2 containing protein complex in β-cell nuclear extracts. TBP-2 binding proteins were purified from INS-1 nuclear extracts by using TBP-2 protein-immobilized (TBP-2) or control beads. ( b ) Identification of the TBP-2 complex. Silver staining was performed. Numbers indicate individual candidate proteins interacting with TBP-2. ( c ) Eluted proteins were analysed by immuno blotting (IB) for Mybbp1a and TBP-2 antibody. ( d ) Co-immunoprecipitation analyses. Input (left, 5% lysate) and anti-Myc immunoprecipitates (right, IP:Myc) from HEK293 cells transfected with pCMV-tag2A and pCMV-tag3B vector (−) or FLAG-HA-Mybbp1a and Myc-TBP-2 vector (+) were analysed by immunoblotting (IB) with antibodies to FLAG, Myc and β-actin. The positions for Mybbp1a (p160; p160MBP) and Mybbp1a (p140; p140MBP) (upper), TBP-2 (middle) and β-actin (lower) are shown. ( e ) Luciferase activity of the UCP-2 -86 enhancer region. INS-1 cells were transfected with PGL4.10 UCP-2 -86 or PGL4.10-luciferase reporter plasmid and each expression plasmid for PGC-1α, Mybbp1a and TBP-2, as indicated. Luciferase reporter activity was normalized by Renilla luciferase activity. ( f ) A schematic model of TBP-2 function in β-cells. Mybbp1a binds PGC-1α (inactive form) and inhibits UCP-2 transcriptional activity. Induced TBP-2 interacts with Mybbp1a and releases PGC-1α from Mybbp1a, facilitating PGC-1α recruitment on the UCP-2 promoter region. Data are presented as mean±s.d. * P

    Article Snippet: Insulin secretion assay using INS-1 cells INS-1 cells were transfected with TBP-2 siRNA (Rn_Txnip_1 and 5) or UCP-2 siRNA (Rn_UCP-2_5 and 6) or negative control siRNA (purchased from Qiagen) for 72 h, or with pCMV-Tag2A-TBP-2 or control pCMV-Tag2A (Stratagene) plasmid for 24 h. Then, the cells were cultured in Krebs Ringer bicarbonate buffer (KRBB) (as mentioned below) or RPMI1640 for INS-1 medium with 2.8 mM (5 mM) or 16.7 mM (20 mM) glucose containing RPMI medium and the supernatant was collected 30 min after the addition of glucose.

    Techniques: Binding Assay, Purification, Silver Staining, Immunoprecipitation, Transfection, Plasmid Preparation, Luciferase, Activity Assay, Expressing, Pyrolysis Gas Chromatography

    Expression of mutant NHS-A proteins in mammalian cells. A : Lysates of HEK 293A cells transiently transfected with GFP-NHS-A400delC and pEGFP-C1 constructs and untransfected cells were analyzed by western blotting with anti-GFP antibody. B : Lysates of HEK 293A cells transiently transfected with FLAG-NHS-A, FLAG-NHS-AC2701T and FLAG-NHS-A4071del299bp in pCMV-Tag 2A and untransfected cells were analyzed by western blotting with anti-FLAG tag antibody. A protein band of greater than 150 kDa seen in all the lanes is due to non-specific binding of the anti-FLAG tag antibody (indicated with an asterisk). A very faint protein band of approximately 130 kDa in the NHS-A and 4071Δ299bp lanes is most likely due to protein degradation. The molecular masses of proteins standards are indicated. UT=untransfected cells.

    Journal: Molecular Vision

    Article Title: Novel causative mutations in patients with Nance-Horan syndrome and altered localization of the mutant NHS-A protein isoform

    doi:

    Figure Lengend Snippet: Expression of mutant NHS-A proteins in mammalian cells. A : Lysates of HEK 293A cells transiently transfected with GFP-NHS-A400delC and pEGFP-C1 constructs and untransfected cells were analyzed by western blotting with anti-GFP antibody. B : Lysates of HEK 293A cells transiently transfected with FLAG-NHS-A, FLAG-NHS-AC2701T and FLAG-NHS-A4071del299bp in pCMV-Tag 2A and untransfected cells were analyzed by western blotting with anti-FLAG tag antibody. A protein band of greater than 150 kDa seen in all the lanes is due to non-specific binding of the anti-FLAG tag antibody (indicated with an asterisk). A very faint protein band of approximately 130 kDa in the NHS-A and 4071Δ299bp lanes is most likely due to protein degradation. The molecular masses of proteins standards are indicated. UT=untransfected cells.

    Article Snippet: The wild type NHS-A cDNA including the exon 3a sequence was cloned in pCMV-Tag 2A (Stratagene, La Jolla, CA) at EcoRI/SalI sites for use as the parent construct for PCR-based mutagenesis.

    Techniques: Expressing, Mutagenesis, Transfection, Construct, Western Blot, FLAG-tag, Binding Assay

    Ectopic expression of TBP-2 in the late stage of HTLV-I-transformed T cells inhibits cell growth and causes apoptosis. I-ED40515T cells ( a ) and I-ATL43T cells ( b ) were transfected with 2 μg pCMV-tag2A-TBP2 or empty vector pCMV-tag2A, respectively. After 48 h, immunoblotting was performed using an anti-Flag antibody to detect Flag-tagged TBP-2 expression (upper panel) and other corresponding antibodies to detect cleaved caspase-3, PARP, β-actin, respectively (middle panel). Flow cytometry assay was performed to detect early apoptotic proportion (lower panel). Data are shown as mean±s.d. ( n =3). *** P

    Journal: Leukemia

    Article Title: Thioredoxin-binding protein-2 (TBP-2/VDUP1/TXNIP) regulates T-cell sensitivity to glucocorticoid during HTLV-I-induced transformation

    doi: 10.1038/leu.2010.286

    Figure Lengend Snippet: Ectopic expression of TBP-2 in the late stage of HTLV-I-transformed T cells inhibits cell growth and causes apoptosis. I-ED40515T cells ( a ) and I-ATL43T cells ( b ) were transfected with 2 μg pCMV-tag2A-TBP2 or empty vector pCMV-tag2A, respectively. After 48 h, immunoblotting was performed using an anti-Flag antibody to detect Flag-tagged TBP-2 expression (upper panel) and other corresponding antibodies to detect cleaved caspase-3, PARP, β-actin, respectively (middle panel). Flow cytometry assay was performed to detect early apoptotic proportion (lower panel). Data are shown as mean±s.d. ( n =3). *** P

    Article Snippet: With regard to the pCMV-tag2A-TBP-2, full-length cDNA of human TBP-2 was cloned in-frame into the pCMV-tag2A vector (Stratagene), as described previously.

    Techniques: Expressing, Transformation Assay, Transfection, Plasmid Preparation, Flow Cytometry, Cytometry