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  • 86
    Thermo Fisher pcdn a3
    ORF 57 increases ORF 59/58 mRNA and protein levels. (A) Top, autoradiogram of a Northern blot containing total RNA from CV-1 cells transfected with a fixed amount of pcDNA 3.1 ORF 59/58 in the presence (+) or absence (−) of 0.1 μg of pcDNA 3.1 ORF 57. A ds DNA probe specific for ORF 59 was used; bottom, the blot was reprobed with a glyceraldehyde-3-phosphate dehydrogenase (GAPDH)-specific probe as a loading control. (B) Western blot analysis of protein extracts from CV-1 cells transfected with a fixed amount of pcDNA 3.1 ORF 59/58 in the presence (+) or absence (−) of 1 μg of pcDNA 3.1 ORF 57. The ORF 59 protein was detected with a KSHV MAb, and then with a secondary goat anti-mouse conjugated to horseradish peroxidase and ECL (Amersham). Equal amounts (10 μg) of extracts as determined by Bradford assay from each transfection were loaded onto the gel.
    Pcdn A3, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Shanghai GenePharma pc dna3 1
    Determination of Ag85A mRNA expression levels by RT-qPCR. Ag85A expression in DC 2.4 cells transfected with <t>pc-DNA3.1,</t> DC2.4 plus pcDNA3.1 (mock-DC) or non-transfected DC 2.4 (Con) for 24 h was determined by RT-qPCR. Data are presented as the mean ± standard error of the mean from three independent tests. **P
    Pc Dna3 1, supplied by Shanghai GenePharma, used in various techniques. Bioz Stars score: 93/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GenePharma Company pc dna3 plasmid
    Determination of Ag85A mRNA expression levels by RT-qPCR. Ag85A expression in DC 2.4 cells transfected with <t>pc-DNA3.1,</t> DC2.4 plus pcDNA3.1 (mock-DC) or non-transfected DC 2.4 (Con) for 24 h was determined by RT-qPCR. Data are presented as the mean ± standard error of the mean from three independent tests. **P
    Pc Dna3 Plasmid, supplied by GenePharma Company, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Genechem pc dna3 1 plasmid
    TGFBR3 interaction with GIPC is critical to the down-regulation of ERK1/2 and JNK signalling by simvastatin. (A) TGFBR3-GIPC complexes were measured by co-IP. (B–E) NMCFs were co-transfected with <t>pc-DNA3.1-m</t> TGFBR3 plasmid and GIPC1 siRNA for 48 h. TGFBR3 and GIPC expression and ERK1/2 and JNK phosphorylation were quantified using Western blot. Data are shown as the means ± SEM. * P
    Pc Dna3 1 Plasmid, supplied by Genechem, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Shanghai Genechem pc dna3 1 plasmid
    Overexpression of TβRIII contributes to the decrease in the viability of CAL-27 cells by TGF-β1 treatment CAL-27 cells were transfected with plasmid encoding TβRIII at concentrations of 0.5 μg/ml and 1 μg/ml. NC represents empty vectors (1 μg/ml of <t>pc-DNA3.1</t> plasmid) transfected into CAL-27 cells, which serves as an NC. After TβRIII overexpression for 24 h, CAL-27 cells were further incubated with 10 ng/ml TGF-β1 for 72 h. ( A ) TβRIII expression as determined by western blot analysis and the averaged band intensities from three independent experiments are presented. ( B ) Relative cell viability as determined by MTT assay. Data is presented as the rate (%) of growth control (viability of control) cultured in growth medium without transfection. Abbreviations: Ctl, control; +NC, empty vector transfection + TGF-β1 48 h; +TβRIII, TβRIII transfection + TGF-β1 48 h. Data are expressed as the mean ± S.D. from three independent experiments ( n =3). * P
    Pc Dna3 1 Plasmid, supplied by Shanghai Genechem, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher pc dna3 1 vector
    Effects of sCLU on E-cadherin mRNA and protein expression. ( A ) Semi-quantitative RT-PCR shows mRNA expression of E-cadherin in HCCLM3 cells treated with increasing concentrations of OGX-011. MM was used as a control in parallel. GAPDH was used as an internal control; ( B ) Semi-quantitative RT-PCR shows mRNA expression of E-cadherin in HepG2 cells transfected with <t>pc.DNA3.1-sCLU</t> for 36 h. pc.DNA3.1 was used as a control in parallel. GAPDH was used as an internal control; ( C ) Western blot analysis shows E-cadherin protein expression in HCCLM3 cells treated with increasing concentrations of OGX-011. MM was used as a control in parallel; ( D ) Western blot analysis shows E-cadherin protein expression in HepG2 cells transfected with pcDNA3.1-sCLU for 36 h. pc.DNA3.1 was used as a control in pa rallel. β-actin was used as an internal control.
    Pc Dna3 1 Vector, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 35 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Genechem pc dna3 1
    Effects of sCLU on E-cadherin mRNA and protein expression. ( A ) Semi-quantitative RT-PCR shows mRNA expression of E-cadherin in HCCLM3 cells treated with increasing concentrations of OGX-011. MM was used as a control in parallel. GAPDH was used as an internal control; ( B ) Semi-quantitative RT-PCR shows mRNA expression of E-cadherin in HepG2 cells transfected with <t>pc.DNA3.1-sCLU</t> for 36 h. pc.DNA3.1 was used as a control in parallel. GAPDH was used as an internal control; ( C ) Western blot analysis shows E-cadherin protein expression in HCCLM3 cells treated with increasing concentrations of OGX-011. MM was used as a control in parallel; ( D ) Western blot analysis shows E-cadherin protein expression in HepG2 cells transfected with pcDNA3.1-sCLU for 36 h. pc.DNA3.1 was used as a control in pa rallel. β-actin was used as an internal control.
    Pc Dna3 1, supplied by Genechem, 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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    88
    Genechem pc dna3 1 mtgfbr3 plasmid
    Effects of sCLU on E-cadherin mRNA and protein expression. ( A ) Semi-quantitative RT-PCR shows mRNA expression of E-cadherin in HCCLM3 cells treated with increasing concentrations of OGX-011. MM was used as a control in parallel. GAPDH was used as an internal control; ( B ) Semi-quantitative RT-PCR shows mRNA expression of E-cadherin in HepG2 cells transfected with <t>pc.DNA3.1-sCLU</t> for 36 h. pc.DNA3.1 was used as a control in parallel. GAPDH was used as an internal control; ( C ) Western blot analysis shows E-cadherin protein expression in HCCLM3 cells treated with increasing concentrations of OGX-011. MM was used as a control in parallel; ( D ) Western blot analysis shows E-cadherin protein expression in HepG2 cells transfected with pcDNA3.1-sCLU for 36 h. pc.DNA3.1 was used as a control in pa rallel. β-actin was used as an internal control.
    Pc Dna3 1 Mtgfbr3 Plasmid, supplied by Genechem, used in various techniques. Bioz Stars score: 88/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Genechem pc dna3 1 mtβriii plasmid
    Effects of sCLU on E-cadherin mRNA and protein expression. ( A ) Semi-quantitative RT-PCR shows mRNA expression of E-cadherin in HCCLM3 cells treated with increasing concentrations of OGX-011. MM was used as a control in parallel. GAPDH was used as an internal control; ( B ) Semi-quantitative RT-PCR shows mRNA expression of E-cadherin in HepG2 cells transfected with <t>pc.DNA3.1-sCLU</t> for 36 h. pc.DNA3.1 was used as a control in parallel. GAPDH was used as an internal control; ( C ) Western blot analysis shows E-cadherin protein expression in HCCLM3 cells treated with increasing concentrations of OGX-011. MM was used as a control in parallel; ( D ) Western blot analysis shows E-cadherin protein expression in HepG2 cells transfected with pcDNA3.1-sCLU for 36 h. pc.DNA3.1 was used as a control in pa rallel. β-actin was used as an internal control.
    Pc Dna3 1 Mtβriii Plasmid, supplied by Genechem, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    GenePharma Company pc dna3 1 ftcd plasmid
    Effects of sCLU on E-cadherin mRNA and protein expression. ( A ) Semi-quantitative RT-PCR shows mRNA expression of E-cadherin in HCCLM3 cells treated with increasing concentrations of OGX-011. MM was used as a control in parallel. GAPDH was used as an internal control; ( B ) Semi-quantitative RT-PCR shows mRNA expression of E-cadherin in HepG2 cells transfected with <t>pc.DNA3.1-sCLU</t> for 36 h. pc.DNA3.1 was used as a control in parallel. GAPDH was used as an internal control; ( C ) Western blot analysis shows E-cadherin protein expression in HCCLM3 cells treated with increasing concentrations of OGX-011. MM was used as a control in parallel; ( D ) Western blot analysis shows E-cadherin protein expression in HepG2 cells transfected with pcDNA3.1-sCLU for 36 h. pc.DNA3.1 was used as a control in pa rallel. β-actin was used as an internal control.
    Pc Dna3 1 Ftcd Plasmid, supplied by GenePharma Company, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Genechem pc dna3 1 hpin1 plasmid
    Effects of sCLU on E-cadherin mRNA and protein expression. ( A ) Semi-quantitative RT-PCR shows mRNA expression of E-cadherin in HCCLM3 cells treated with increasing concentrations of OGX-011. MM was used as a control in parallel. GAPDH was used as an internal control; ( B ) Semi-quantitative RT-PCR shows mRNA expression of E-cadherin in HepG2 cells transfected with <t>pc.DNA3.1-sCLU</t> for 36 h. pc.DNA3.1 was used as a control in parallel. GAPDH was used as an internal control; ( C ) Western blot analysis shows E-cadherin protein expression in HCCLM3 cells treated with increasing concentrations of OGX-011. MM was used as a control in parallel; ( D ) Western blot analysis shows E-cadherin protein expression in HepG2 cells transfected with pcDNA3.1-sCLU for 36 h. pc.DNA3.1 was used as a control in pa rallel. β-actin was used as an internal control.
    Pc Dna3 1 Hpin1 Plasmid, supplied by Genechem, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Genechem pc dna3 1 htgfbr3 plasmid
    Effects of sCLU on E-cadherin mRNA and protein expression. ( A ) Semi-quantitative RT-PCR shows mRNA expression of E-cadherin in HCCLM3 cells treated with increasing concentrations of OGX-011. MM was used as a control in parallel. GAPDH was used as an internal control; ( B ) Semi-quantitative RT-PCR shows mRNA expression of E-cadherin in HepG2 cells transfected with <t>pc.DNA3.1-sCLU</t> for 36 h. pc.DNA3.1 was used as a control in parallel. GAPDH was used as an internal control; ( C ) Western blot analysis shows E-cadherin protein expression in HCCLM3 cells treated with increasing concentrations of OGX-011. MM was used as a control in parallel; ( D ) Western blot analysis shows E-cadherin protein expression in HepG2 cells transfected with pcDNA3.1-sCLU for 36 h. pc.DNA3.1 was used as a control in pa rallel. β-actin was used as an internal control.
    Pc Dna3 1 Htgfbr3 Plasmid, supplied by Genechem, used in various techniques. Bioz Stars score: 91/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Addgene inc 5446 bp plasmid pc dna3 egfp
    Effects of sCLU on E-cadherin mRNA and protein expression. ( A ) Semi-quantitative RT-PCR shows mRNA expression of E-cadherin in HCCLM3 cells treated with increasing concentrations of OGX-011. MM was used as a control in parallel. GAPDH was used as an internal control; ( B ) Semi-quantitative RT-PCR shows mRNA expression of E-cadherin in HepG2 cells transfected with <t>pc.DNA3.1-sCLU</t> for 36 h. pc.DNA3.1 was used as a control in parallel. GAPDH was used as an internal control; ( C ) Western blot analysis shows E-cadherin protein expression in HCCLM3 cells treated with increasing concentrations of OGX-011. MM was used as a control in parallel; ( D ) Western blot analysis shows E-cadherin protein expression in HepG2 cells transfected with pcDNA3.1-sCLU for 36 h. pc.DNA3.1 was used as a control in pa rallel. β-actin was used as an internal control.
    5446 Bp Plasmid Pc Dna3 Egfp, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Addgene inc plasmid pc dna3 flag lkb1
    Characterization of HeLa cells expressing <t>LKB1.</t> (A) HeLa cells were transfected with a plasmid encoding LKB1 <t>(pc-DNA3-flag-LKB1)</t> or vector and were subjected to G418 selection for two weeks. LKB1 was detected by western blot. GAPDH was used as a loading
    Plasmid Pc Dna3 Flag Lkb1, supplied by Addgene inc, used in various techniques. Bioz Stars score: 86/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher pc dna3 1 myc
    Characterization of HeLa cells expressing <t>LKB1.</t> (A) HeLa cells were transfected with a plasmid encoding LKB1 <t>(pc-DNA3-flag-LKB1)</t> or vector and were subjected to G418 selection for two weeks. LKB1 was detected by western blot. GAPDH was used as a loading
    Pc Dna3 1 Myc, supplied by Thermo Fisher, 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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    Thermo Fisher pc dna3 1 flag vectors
    Characterization of HeLa cells expressing <t>LKB1.</t> (A) HeLa cells were transfected with a plasmid encoding LKB1 <t>(pc-DNA3-flag-LKB1)</t> or vector and were subjected to G418 selection for two weeks. LKB1 was detected by western blot. GAPDH was used as a loading
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    Genechem reagents pc dna3 1 mtgfbr3 plasmid
    Characterization of HeLa cells expressing <t>LKB1.</t> (A) HeLa cells were transfected with a plasmid encoding LKB1 <t>(pc-DNA3-flag-LKB1)</t> or vector and were subjected to G418 selection for two weeks. LKB1 was detected by western blot. GAPDH was used as a loading
    Reagents Pc Dna3 1 Mtgfbr3 Plasmid, supplied by Genechem, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa expression vector pc dna3 1
    Characterization of HeLa cells expressing <t>LKB1.</t> (A) HeLa cells were transfected with a plasmid encoding LKB1 <t>(pc-DNA3-flag-LKB1)</t> or vector and were subjected to G418 selection for two weeks. LKB1 was detected by western blot. GAPDH was used as a loading
    Expression Vector Pc Dna3 1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa pcdna 3
    Characterization of HeLa cells expressing <t>LKB1.</t> (A) HeLa cells were transfected with a plasmid encoding LKB1 <t>(pc-DNA3-flag-LKB1)</t> or vector and were subjected to G418 selection for two weeks. LKB1 was detected by western blot. GAPDH was used as a loading
    Pcdna 3, supplied by TaKaRa, used in various techniques. Bioz Stars score: 90/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega pcdna 3
    Characterization of HeLa cells expressing <t>LKB1.</t> (A) HeLa cells were transfected with a plasmid encoding LKB1 <t>(pc-DNA3-flag-LKB1)</t> or vector and were subjected to G418 selection for two weeks. LKB1 was detected by western blot. GAPDH was used as a loading
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    Image Search Results


    ORF 57 increases ORF 59/58 mRNA and protein levels. (A) Top, autoradiogram of a Northern blot containing total RNA from CV-1 cells transfected with a fixed amount of pcDNA 3.1 ORF 59/58 in the presence (+) or absence (−) of 0.1 μg of pcDNA 3.1 ORF 57. A ds DNA probe specific for ORF 59 was used; bottom, the blot was reprobed with a glyceraldehyde-3-phosphate dehydrogenase (GAPDH)-specific probe as a loading control. (B) Western blot analysis of protein extracts from CV-1 cells transfected with a fixed amount of pcDNA 3.1 ORF 59/58 in the presence (+) or absence (−) of 1 μg of pcDNA 3.1 ORF 57. The ORF 59 protein was detected with a KSHV MAb, and then with a secondary goat anti-mouse conjugated to horseradish peroxidase and ECL (Amersham). Equal amounts (10 μg) of extracts as determined by Bradford assay from each transfection were loaded onto the gel.

    Journal: Journal of Virology

    Article Title: Kaposi's Sarcoma-Associated Herpesvirus Open Reading Frame 57 Encodes a Posttranscriptional Regulator with Multiple Distinct Activities †

    doi:

    Figure Lengend Snippet: ORF 57 increases ORF 59/58 mRNA and protein levels. (A) Top, autoradiogram of a Northern blot containing total RNA from CV-1 cells transfected with a fixed amount of pcDNA 3.1 ORF 59/58 in the presence (+) or absence (−) of 0.1 μg of pcDNA 3.1 ORF 57. A ds DNA probe specific for ORF 59 was used; bottom, the blot was reprobed with a glyceraldehyde-3-phosphate dehydrogenase (GAPDH)-specific probe as a loading control. (B) Western blot analysis of protein extracts from CV-1 cells transfected with a fixed amount of pcDNA 3.1 ORF 59/58 in the presence (+) or absence (−) of 1 μg of pcDNA 3.1 ORF 57. The ORF 59 protein was detected with a KSHV MAb, and then with a secondary goat anti-mouse conjugated to horseradish peroxidase and ECL (Amersham). Equal amounts (10 μg) of extracts as determined by Bradford assay from each transfection were loaded onto the gel.

    Article Snippet: The resulting fragment was cloned into the Hin dIII and Eco RV sites of pcDNA 3 (Invitrogen) to create pcDNA 3 nut-1.

    Techniques: Northern Blot, Transfection, Western Blot, Bradford Assay

    ). Forty-eight hours posttransfection, luciferase activity was measured and values were plotted that represented fold activation relative to that of ORF 50 alone, which was set to 1. (C) Mutations in the nut-1 promoter abolish synergy between ORF 50 and ORF 57. CV-1 cells were transfected with 2 μg of pcDNA 3 ORF 50, increasing amounts of ORF 57, and either 1 μg of the wild-type nut-1-706 reporter or the nut-1-706 mutant reporter. Forty-eight hours posttransfection, luciferase activity was measured and values were plotted that represent fold activation relative to that of ORF 50 alone, which was set to 1.

    Journal: Journal of Virology

    Article Title: Kaposi's Sarcoma-Associated Herpesvirus Open Reading Frame 57 Encodes a Posttranscriptional Regulator with Multiple Distinct Activities †

    doi:

    Figure Lengend Snippet: ). Forty-eight hours posttransfection, luciferase activity was measured and values were plotted that represented fold activation relative to that of ORF 50 alone, which was set to 1. (C) Mutations in the nut-1 promoter abolish synergy between ORF 50 and ORF 57. CV-1 cells were transfected with 2 μg of pcDNA 3 ORF 50, increasing amounts of ORF 57, and either 1 μg of the wild-type nut-1-706 reporter or the nut-1-706 mutant reporter. Forty-eight hours posttransfection, luciferase activity was measured and values were plotted that represent fold activation relative to that of ORF 50 alone, which was set to 1.

    Article Snippet: The resulting fragment was cloned into the Hin dIII and Eco RV sites of pcDNA 3 (Invitrogen) to create pcDNA 3 nut-1.

    Techniques: Luciferase, Activity Assay, Activation Assay, Transfection, Mutagenesis

    The C-terminally tagged version of ORF 57 is localized to the nucleus. The panels show results of immunofluorescence staining of CV-1 cells 48 h after transfection with either empty pcDNA 3.1-V5 vector (left panel) or pcDNA 3.1-V5 ORF 57 (right panel). Reactivity to anti-V5 antibody (Invitrogen) was detected with a TRITC-conjugated rabbit anti-mouse antibody.

    Journal: Journal of Virology

    Article Title: Kaposi's Sarcoma-Associated Herpesvirus Open Reading Frame 57 Encodes a Posttranscriptional Regulator with Multiple Distinct Activities †

    doi:

    Figure Lengend Snippet: The C-terminally tagged version of ORF 57 is localized to the nucleus. The panels show results of immunofluorescence staining of CV-1 cells 48 h after transfection with either empty pcDNA 3.1-V5 vector (left panel) or pcDNA 3.1-V5 ORF 57 (right panel). Reactivity to anti-V5 antibody (Invitrogen) was detected with a TRITC-conjugated rabbit anti-mouse antibody.

    Article Snippet: The resulting fragment was cloned into the Hin dIII and Eco RV sites of pcDNA 3 (Invitrogen) to create pcDNA 3 nut-1.

    Techniques: Immunofluorescence, Staining, Transfection, Plasmid Preparation

    ORF 50 mRNA and protein levels are not significantly increased by ORF 57 expression. (A) Top, autoradiogram of a Northern blot containing total RNA from CV-1 cells transfected with a fixed amount of pcDNA 3 ORF 50 and the following amounts of pcDNA 3.1 ORF 57: 0 μg (lane 1), 0.75 μg (lane 2), 2.5 μg (lane 3), and 7.5 μg (lane 4). An antisense riboprobe for ORF 50 was hybridized to the blot. Bottom, the blot was reprobed with a GAPDH-specific probe as a loading control. (B) Western blot analysis of protein extracts from CV-1 cells transfected with a fixed amount of pcDNA 3.1 ORF 50 and the following amounts of pcDNA 3.1 ORF 57: 0 μg (lane 1), 0.5 μg (lane 2), 2.5 μg (lane 3), and 5 μg (lane 4). The ORF 50 protein was detected with a polyclonal antibody and then with a secondary rabbit anti-mouse antibody conjugated to horseradish peroxidase and ECL (Amersham). Equal amount of extracts as determined by Bradford assay from each transfection were loaded onto the gel.

    Journal: Journal of Virology

    Article Title: Kaposi's Sarcoma-Associated Herpesvirus Open Reading Frame 57 Encodes a Posttranscriptional Regulator with Multiple Distinct Activities †

    doi:

    Figure Lengend Snippet: ORF 50 mRNA and protein levels are not significantly increased by ORF 57 expression. (A) Top, autoradiogram of a Northern blot containing total RNA from CV-1 cells transfected with a fixed amount of pcDNA 3 ORF 50 and the following amounts of pcDNA 3.1 ORF 57: 0 μg (lane 1), 0.75 μg (lane 2), 2.5 μg (lane 3), and 7.5 μg (lane 4). An antisense riboprobe for ORF 50 was hybridized to the blot. Bottom, the blot was reprobed with a GAPDH-specific probe as a loading control. (B) Western blot analysis of protein extracts from CV-1 cells transfected with a fixed amount of pcDNA 3.1 ORF 50 and the following amounts of pcDNA 3.1 ORF 57: 0 μg (lane 1), 0.5 μg (lane 2), 2.5 μg (lane 3), and 5 μg (lane 4). The ORF 50 protein was detected with a polyclonal antibody and then with a secondary rabbit anti-mouse antibody conjugated to horseradish peroxidase and ECL (Amersham). Equal amount of extracts as determined by Bradford assay from each transfection were loaded onto the gel.

    Article Snippet: The resulting fragment was cloned into the Hin dIII and Eco RV sites of pcDNA 3 (Invitrogen) to create pcDNA 3 nut-1.

    Techniques: Expressing, Northern Blot, Transfection, Western Blot, Bradford Assay

    Effect of introns on synergistic activation of different promoters by the ORF 50-ORF 57 combination. CV-1 cells were cotransfected with fixed amounts of the indicated KSHV promoter constructs with or without introns (Nut-1, TK, and Kaposin), a fixed amount of the transcriptional activator, pcDNA 3 ORF 50, and increasing amounts of pcDNA 3.1 ORF 57 (0, 0.3, 1, and 3 μg). Luciferase activity was measured 48 h after transfection. (A) Log scale graph displaying the effects on the Nut-1 promoter with and without an intron. (B) Graph displaying the effects on the Kaposin and TK promoters with and without an intron. Values are plotted as fold luciferase activity over that of ORF 50 alone, which was set to 1. Error bars representing standard deviations are from two experiments performed in duplicate. Note that activation is plotted on an arithmetic scale in B and on a log scale in A.

    Journal: Journal of Virology

    Article Title: Kaposi's Sarcoma-Associated Herpesvirus Open Reading Frame 57 Encodes a Posttranscriptional Regulator with Multiple Distinct Activities †

    doi:

    Figure Lengend Snippet: Effect of introns on synergistic activation of different promoters by the ORF 50-ORF 57 combination. CV-1 cells were cotransfected with fixed amounts of the indicated KSHV promoter constructs with or without introns (Nut-1, TK, and Kaposin), a fixed amount of the transcriptional activator, pcDNA 3 ORF 50, and increasing amounts of pcDNA 3.1 ORF 57 (0, 0.3, 1, and 3 μg). Luciferase activity was measured 48 h after transfection. (A) Log scale graph displaying the effects on the Nut-1 promoter with and without an intron. (B) Graph displaying the effects on the Kaposin and TK promoters with and without an intron. Values are plotted as fold luciferase activity over that of ORF 50 alone, which was set to 1. Error bars representing standard deviations are from two experiments performed in duplicate. Note that activation is plotted on an arithmetic scale in B and on a log scale in A.

    Article Snippet: The resulting fragment was cloned into the Hin dIII and Eco RV sites of pcDNA 3 (Invitrogen) to create pcDNA 3 nut-1.

    Techniques: Activation Assay, Construct, Luciferase, Activity Assay, Transfection

    Effect of ORF 57 expression on viral promoter constructs with and without introns. CV-1 cells were cotransfected with increasing amounts of pcDNA 3.1 ORF 57 (0, 0.3, 1, and 3 μg) and fixed amounts of the following KSHV promoter luciferase constructs with or without an intron: DNA Pol, Kaposin, Nut-1, and TK. Luciferase activity was measured 48 h posttransfection. Graphs are representative examples of experiments performed in duplicate. Error bars represent standard deviations.

    Journal: Journal of Virology

    Article Title: Kaposi's Sarcoma-Associated Herpesvirus Open Reading Frame 57 Encodes a Posttranscriptional Regulator with Multiple Distinct Activities †

    doi:

    Figure Lengend Snippet: Effect of ORF 57 expression on viral promoter constructs with and without introns. CV-1 cells were cotransfected with increasing amounts of pcDNA 3.1 ORF 57 (0, 0.3, 1, and 3 μg) and fixed amounts of the following KSHV promoter luciferase constructs with or without an intron: DNA Pol, Kaposin, Nut-1, and TK. Luciferase activity was measured 48 h posttransfection. Graphs are representative examples of experiments performed in duplicate. Error bars represent standard deviations.

    Article Snippet: The resulting fragment was cloned into the Hin dIII and Eco RV sites of pcDNA 3 (Invitrogen) to create pcDNA 3 nut-1.

    Techniques: Expressing, Construct, Luciferase, Activity Assay

    Upregulation of nut-1 RNA accumulation by ORF 57 expression. The indicated constructs (bottom) were transfected into CV-1 cells in the presence (+) or absence (−) of 1 μg of pcDNA 3.1 ORF 57. Forty-eight hours later, total RNA was prepared from whole-cell lysates, electrophoresed through 1% agarose-formaldehyde gels, transferred to Hybond-N+ membranes, and hybridized to probes specific for nut-1 (A) or ORF 74/GCR (B, left panel) or ORF K5 (B, right panel). Below each panel is shown rRNA bands in ethidium bromide-stained lanes as a loading control.

    Journal: Journal of Virology

    Article Title: Kaposi's Sarcoma-Associated Herpesvirus Open Reading Frame 57 Encodes a Posttranscriptional Regulator with Multiple Distinct Activities †

    doi:

    Figure Lengend Snippet: Upregulation of nut-1 RNA accumulation by ORF 57 expression. The indicated constructs (bottom) were transfected into CV-1 cells in the presence (+) or absence (−) of 1 μg of pcDNA 3.1 ORF 57. Forty-eight hours later, total RNA was prepared from whole-cell lysates, electrophoresed through 1% agarose-formaldehyde gels, transferred to Hybond-N+ membranes, and hybridized to probes specific for nut-1 (A) or ORF 74/GCR (B, left panel) or ORF K5 (B, right panel). Below each panel is shown rRNA bands in ethidium bromide-stained lanes as a loading control.

    Article Snippet: The resulting fragment was cloned into the Hin dIII and Eco RV sites of pcDNA 3 (Invitrogen) to create pcDNA 3 nut-1.

    Techniques: Expressing, Construct, Transfection, Staining

    Determination of Ag85A mRNA expression levels by RT-qPCR. Ag85A expression in DC 2.4 cells transfected with pc-DNA3.1, DC2.4 plus pcDNA3.1 (mock-DC) or non-transfected DC 2.4 (Con) for 24 h was determined by RT-qPCR. Data are presented as the mean ± standard error of the mean from three independent tests. **P

    Journal: Experimental and Therapeutic Medicine

    Article Title: Dendritic cell vaccine with Ag85A enhances anti-colorectal carcinoma immunity

    doi: 10.3892/etm.2018.6851

    Figure Lengend Snippet: Determination of Ag85A mRNA expression levels by RT-qPCR. Ag85A expression in DC 2.4 cells transfected with pc-DNA3.1, DC2.4 plus pcDNA3.1 (mock-DC) or non-transfected DC 2.4 (Con) for 24 h was determined by RT-qPCR. Data are presented as the mean ± standard error of the mean from three independent tests. **P

    Article Snippet: Ag85A plasmid transfection Cells were transfected with 1 µg Ag85A and pc-DNA3.1 (Shanghai GenePharma Co., Ltd., Shanghai, China) using Lipofectamine 2000 transfection reagent (Invitrogen; Thermo Fisher Scientific, Inc.).

    Techniques: Expressing, Quantitative RT-PCR, Transfection

    TGFBR3 interaction with GIPC is critical to the down-regulation of ERK1/2 and JNK signalling by simvastatin. (A) TGFBR3-GIPC complexes were measured by co-IP. (B–E) NMCFs were co-transfected with pc-DNA3.1-m TGFBR3 plasmid and GIPC1 siRNA for 48 h. TGFBR3 and GIPC expression and ERK1/2 and JNK phosphorylation were quantified using Western blot. Data are shown as the means ± SEM. * P

    Journal: British Journal of Pharmacology

    Article Title: Simvastatin alleviates cardiac fibrosis induced by infarction via up-regulation of TGF-β receptor III expression

    doi: 10.1111/bph.13166

    Figure Lengend Snippet: TGFBR3 interaction with GIPC is critical to the down-regulation of ERK1/2 and JNK signalling by simvastatin. (A) TGFBR3-GIPC complexes were measured by co-IP. (B–E) NMCFs were co-transfected with pc-DNA3.1-m TGFBR3 plasmid and GIPC1 siRNA for 48 h. TGFBR3 and GIPC expression and ERK1/2 and JNK phosphorylation were quantified using Western blot. Data are shown as the means ± SEM. * P

    Article Snippet: Pc-DNA3.1-mTGFBR3 plasmid (GeneChem Co., Ltd, Shanghai, China) and pc-DNA3.1-plasmid (GeneChem Co., Ltd) as an empty vector were all transfected with X-tremeGENE siRNA transfection reagent (Roche Molecular Biochemicals, Mannheim, Germany) for 48 h.

    Techniques: Co-Immunoprecipitation Assay, Transfection, Plasmid Preparation, Expressing, Western Blot

    Overexpression of TβRIII contributes to the decrease in the viability of CAL-27 cells by TGF-β1 treatment CAL-27 cells were transfected with plasmid encoding TβRIII at concentrations of 0.5 μg/ml and 1 μg/ml. NC represents empty vectors (1 μg/ml of pc-DNA3.1 plasmid) transfected into CAL-27 cells, which serves as an NC. After TβRIII overexpression for 24 h, CAL-27 cells were further incubated with 10 ng/ml TGF-β1 for 72 h. ( A ) TβRIII expression as determined by western blot analysis and the averaged band intensities from three independent experiments are presented. ( B ) Relative cell viability as determined by MTT assay. Data is presented as the rate (%) of growth control (viability of control) cultured in growth medium without transfection. Abbreviations: Ctl, control; +NC, empty vector transfection + TGF-β1 48 h; +TβRIII, TβRIII transfection + TGF-β1 48 h. Data are expressed as the mean ± S.D. from three independent experiments ( n =3). * P

    Journal: Bioscience Reports

    Article Title: Overexpression of transforming growth factor type III receptor restores TGF-β1 sensitivity in human tongue squamous cell carcinoma cells

    doi: 10.1042/BSR20150141

    Figure Lengend Snippet: Overexpression of TβRIII contributes to the decrease in the viability of CAL-27 cells by TGF-β1 treatment CAL-27 cells were transfected with plasmid encoding TβRIII at concentrations of 0.5 μg/ml and 1 μg/ml. NC represents empty vectors (1 μg/ml of pc-DNA3.1 plasmid) transfected into CAL-27 cells, which serves as an NC. After TβRIII overexpression for 24 h, CAL-27 cells were further incubated with 10 ng/ml TGF-β1 for 72 h. ( A ) TβRIII expression as determined by western blot analysis and the averaged band intensities from three independent experiments are presented. ( B ) Relative cell viability as determined by MTT assay. Data is presented as the rate (%) of growth control (viability of control) cultured in growth medium without transfection. Abbreviations: Ctl, control; +NC, empty vector transfection + TGF-β1 48 h; +TβRIII, TβRIII transfection + TGF-β1 48 h. Data are expressed as the mean ± S.D. from three independent experiments ( n =3). * P

    Article Snippet: The pc-DNA3.1-plasmid (Shanghai GeneChem Co.) was used as an empty vector and Lipofectamine® 2000 was used as the transfection reagent.

    Techniques: Over Expression, Transfection, Plasmid Preparation, Incubation, Expressing, Western Blot, MTT Assay, Cell Culture, CTL Assay

    Effects of sCLU on E-cadherin mRNA and protein expression. ( A ) Semi-quantitative RT-PCR shows mRNA expression of E-cadherin in HCCLM3 cells treated with increasing concentrations of OGX-011. MM was used as a control in parallel. GAPDH was used as an internal control; ( B ) Semi-quantitative RT-PCR shows mRNA expression of E-cadherin in HepG2 cells transfected with pc.DNA3.1-sCLU for 36 h. pc.DNA3.1 was used as a control in parallel. GAPDH was used as an internal control; ( C ) Western blot analysis shows E-cadherin protein expression in HCCLM3 cells treated with increasing concentrations of OGX-011. MM was used as a control in parallel; ( D ) Western blot analysis shows E-cadherin protein expression in HepG2 cells transfected with pcDNA3.1-sCLU for 36 h. pc.DNA3.1 was used as a control in pa rallel. β-actin was used as an internal control.

    Journal: International Journal of Molecular Sciences

    Article Title: Antisense Oligonucleotide Against Clusterin Regulates Human Hepatocellular Carcinoma Invasion Through Transcriptional Regulation of Matrix Metalloproteinase-2 and E-Cadherin

    doi: 10.3390/ijms130810594

    Figure Lengend Snippet: Effects of sCLU on E-cadherin mRNA and protein expression. ( A ) Semi-quantitative RT-PCR shows mRNA expression of E-cadherin in HCCLM3 cells treated with increasing concentrations of OGX-011. MM was used as a control in parallel. GAPDH was used as an internal control; ( B ) Semi-quantitative RT-PCR shows mRNA expression of E-cadherin in HepG2 cells transfected with pc.DNA3.1-sCLU for 36 h. pc.DNA3.1 was used as a control in parallel. GAPDH was used as an internal control; ( C ) Western blot analysis shows E-cadherin protein expression in HCCLM3 cells treated with increasing concentrations of OGX-011. MM was used as a control in parallel; ( D ) Western blot analysis shows E-cadherin protein expression in HepG2 cells transfected with pcDNA3.1-sCLU for 36 h. pc.DNA3.1 was used as a control in pa rallel. β-actin was used as an internal control.

    Article Snippet: From this vector, clusterin full-length cDNA was then subcloned into the pc.DNA3.1 vector (Invitrogen, Carlsbad, CA) that was previously digested with Hin dIII and treated with calf intestinal alkaline phosphatase to produce pc.DNA3.1-sCLU.

    Techniques: Expressing, Quantitative RT-PCR, Transfection, Western Blot

    Effect of sCLU overexpression on MMP-2 expression and activity. ( A ) HepG2 cells were transfected with pc.DNA3.1-sCLU or pcDNA3.1 for 36 h. Representative images showing expression of MMP-2 mRNA in transfected cells, as determined by RT-PCR; ( B ) Western blot analysis to evaluate pro-MMP-2 protein expression in pc.DNA3.1-sCLU or pc.DNA3.1-transfected HepG2 cells. Blot was reprobed with β-actin antibody to verify the equal loading of proteins; ( C ) Gelatin zymogram showing activity of pro-MMP-2 and active MMP-2 in pc.DNA3.1-sCLU or pc.DNA3.1-transfected HepG2 cells. Columns, mean of quadruple experiments; bars, SD. * p

    Journal: International Journal of Molecular Sciences

    Article Title: Antisense Oligonucleotide Against Clusterin Regulates Human Hepatocellular Carcinoma Invasion Through Transcriptional Regulation of Matrix Metalloproteinase-2 and E-Cadherin

    doi: 10.3390/ijms130810594

    Figure Lengend Snippet: Effect of sCLU overexpression on MMP-2 expression and activity. ( A ) HepG2 cells were transfected with pc.DNA3.1-sCLU or pcDNA3.1 for 36 h. Representative images showing expression of MMP-2 mRNA in transfected cells, as determined by RT-PCR; ( B ) Western blot analysis to evaluate pro-MMP-2 protein expression in pc.DNA3.1-sCLU or pc.DNA3.1-transfected HepG2 cells. Blot was reprobed with β-actin antibody to verify the equal loading of proteins; ( C ) Gelatin zymogram showing activity of pro-MMP-2 and active MMP-2 in pc.DNA3.1-sCLU or pc.DNA3.1-transfected HepG2 cells. Columns, mean of quadruple experiments; bars, SD. * p

    Article Snippet: From this vector, clusterin full-length cDNA was then subcloned into the pc.DNA3.1 vector (Invitrogen, Carlsbad, CA) that was previously digested with Hin dIII and treated with calf intestinal alkaline phosphatase to produce pc.DNA3.1-sCLU.

    Techniques: Over Expression, Expressing, Activity Assay, Transfection, Reverse Transcription Polymerase Chain Reaction, Western Blot

    Effect of sCLU overexpression on the invasive capability of HepG2 cells. ( A ) Western blot analysis of sCLU expression in cells transfected with pc.DNA3.1 (vector) or pc.DNA3.1-sCLU. Histogram represents the relative density of sCLU bands normalized to β-actin; ( B ) Histogram showing the invasive capability of transfected HepG2 cells. The experiment was done in triplicate and the value obtained from pc.DNA3.1 transfected cells was set at 100%. Each bar represents mean ± SE ( n = 3); * p

    Journal: International Journal of Molecular Sciences

    Article Title: Antisense Oligonucleotide Against Clusterin Regulates Human Hepatocellular Carcinoma Invasion Through Transcriptional Regulation of Matrix Metalloproteinase-2 and E-Cadherin

    doi: 10.3390/ijms130810594

    Figure Lengend Snippet: Effect of sCLU overexpression on the invasive capability of HepG2 cells. ( A ) Western blot analysis of sCLU expression in cells transfected with pc.DNA3.1 (vector) or pc.DNA3.1-sCLU. Histogram represents the relative density of sCLU bands normalized to β-actin; ( B ) Histogram showing the invasive capability of transfected HepG2 cells. The experiment was done in triplicate and the value obtained from pc.DNA3.1 transfected cells was set at 100%. Each bar represents mean ± SE ( n = 3); * p

    Article Snippet: From this vector, clusterin full-length cDNA was then subcloned into the pc.DNA3.1 vector (Invitrogen, Carlsbad, CA) that was previously digested with Hin dIII and treated with calf intestinal alkaline phosphatase to produce pc.DNA3.1-sCLU.

    Techniques: Over Expression, Western Blot, Expressing, Transfection, Plasmid Preparation

    Characterization of HeLa cells expressing LKB1. (A) HeLa cells were transfected with a plasmid encoding LKB1 (pc-DNA3-flag-LKB1) or vector and were subjected to G418 selection for two weeks. LKB1 was detected by western blot. GAPDH was used as a loading

    Journal: Gynecologic oncology

    Article Title: Expression and transcriptional profiling of the LKB1 tumor suppressor in cervical cancer cells

    doi: 10.1016/j.ygyno.2014.04.050

    Figure Lengend Snippet: Characterization of HeLa cells expressing LKB1. (A) HeLa cells were transfected with a plasmid encoding LKB1 (pc-DNA3-flag-LKB1) or vector and were subjected to G418 selection for two weeks. LKB1 was detected by western blot. GAPDH was used as a loading

    Article Snippet: The plasmid pc-DNA3-flag-LKB1 was bought from Addgene (Cambridge, MA).

    Techniques: Expressing, Transfection, Plasmid Preparation, Selection, Western Blot