vhl cdna vectors Search Results


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  • 99
    Qiagen expression vector pqe 30
    Expression Vector Pqe 30, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 380 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 380 article reviews
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    expression vector pqe 30 - by Bioz Stars, 2020-04
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    99
    Thermo Fisher pef6 myc his a
    Pef6 Myc His A, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 61 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    pef6 myc his a - by Bioz Stars, 2020-04
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    87
    OriGene hif 1α cdna
    EGFR -activating mutations are associated with elevated MET and <t>HIF-1α</t> levels in NSCLC cell lines. ( a ) Gene expression analysis was performed on gene arrays of 53 NSCLC lines. MET expression was elevated in NSCLC cell lines harboring EGFR -activating mutations; * P = 0.002. ( b ) MET expression in 53 NSCLC cell lines with high EGFR gene copy number ( > 4 copies) vs low copy number (
    Hif 1α Cdna, supplied by OriGene, used in various techniques. Bioz Stars score: 87/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher cdnas
    EGFR -activating mutations are associated with elevated MET and <t>HIF-1α</t> levels in NSCLC cell lines. ( a ) Gene expression analysis was performed on gene arrays of 53 NSCLC lines. MET expression was elevated in NSCLC cell lines harboring EGFR -activating mutations; * P = 0.002. ( b ) MET expression in 53 NSCLC cell lines with high EGFR gene copy number ( > 4 copies) vs low copy number (
    Cdnas, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 53579 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    cdnas - by Bioz Stars, 2020-04
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    93
    Stratagene cdnas
    EGFR -activating mutations are associated with elevated MET and <t>HIF-1α</t> levels in NSCLC cell lines. ( a ) Gene expression analysis was performed on gene arrays of 53 NSCLC lines. MET expression was elevated in NSCLC cell lines harboring EGFR -activating mutations; * P = 0.002. ( b ) MET expression in 53 NSCLC cell lines with high EGFR gene copy number ( > 4 copies) vs low copy number (
    Cdnas, supplied by Stratagene, used in various techniques. Bioz Stars score: 93/100, based on 3580 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    TaKaRa human kidney cdna library
    EGFR -activating mutations are associated with elevated MET and <t>HIF-1α</t> levels in NSCLC cell lines. ( a ) Gene expression analysis was performed on gene arrays of 53 NSCLC lines. MET expression was elevated in NSCLC cell lines harboring EGFR -activating mutations; * P = 0.002. ( b ) MET expression in 53 NSCLC cell lines with high EGFR gene copy number ( > 4 copies) vs low copy number (
    Human Kidney Cdna Library, supplied by TaKaRa, used in various techniques. Bioz Stars score: 93/100, based on 299 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    87
    TaKaRa pretransformed human kidney cdna library
    EGFR -activating mutations are associated with elevated MET and <t>HIF-1α</t> levels in NSCLC cell lines. ( a ) Gene expression analysis was performed on gene arrays of 53 NSCLC lines. MET expression was elevated in NSCLC cell lines harboring EGFR -activating mutations; * P = 0.002. ( b ) MET expression in 53 NSCLC cell lines with high EGFR gene copy number ( > 4 copies) vs low copy number (
    Pretransformed Human Kidney Cdna Library, supplied by TaKaRa, used in various techniques. Bioz Stars score: 87/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Addgene inc stable cells expressing keap1 constructs pbabe parental vector
    EGFR -activating mutations are associated with elevated MET and <t>HIF-1α</t> levels in NSCLC cell lines. ( a ) Gene expression analysis was performed on gene arrays of 53 NSCLC lines. MET expression was elevated in NSCLC cell lines harboring EGFR -activating mutations; * P = 0.002. ( b ) MET expression in 53 NSCLC cell lines with high EGFR gene copy number ( > 4 copies) vs low copy number (
    Stable Cells Expressing Keap1 Constructs Pbabe Parental Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    87
    Addgene inc stable cells expressing keap1 mutant constructs pbabe parental vector
    EGFR -activating mutations are associated with elevated MET and <t>HIF-1α</t> levels in NSCLC cell lines. ( a ) Gene expression analysis was performed on gene arrays of 53 NSCLC lines. MET expression was elevated in NSCLC cell lines harboring EGFR -activating mutations; * P = 0.002. ( b ) MET expression in 53 NSCLC cell lines with high EGFR gene copy number ( > 4 copies) vs low copy number (
    Stable Cells Expressing Keap1 Mutant Constructs Pbabe Parental Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 87/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Addgene inc retroviral expression vector pbabe puro
    EGFR -activating mutations are associated with elevated MET and <t>HIF-1α</t> levels in NSCLC cell lines. ( a ) Gene expression analysis was performed on gene arrays of 53 NSCLC lines. MET expression was elevated in NSCLC cell lines harboring EGFR -activating mutations; * P = 0.002. ( b ) MET expression in 53 NSCLC cell lines with high EGFR gene copy number ( > 4 copies) vs low copy number (
    Retroviral Expression Vector Pbabe Puro, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 52 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    TaKaRa pretrox ptight tet on vectors
    Degradation of VPS34 through AdPROM results in loss of function. ( a ) GFP-VPS34 knockin HEK293 cells expressing the <t>Tet-transactivator</t> were infected with <t>pRetroX-Tight</t> empty vector control. Following selection, cells were seeded onto glass coverslips and either left untreated or treated with doxycycline (2 µg ml −1 ; 24 h) or VPS34-IN1 (2.5 µM, 1 h). Cells were permealized in liquid nitrogen, and fixed in 3.7% (w/v) paraformaldehyde before staining with fluorescently labelled selective PI3P binding (green) and interaction deficient (red) probes as described in the Material and methods section. Samples were mounted on microscopy slides with mounting media containing DAPI (blue). ( b ) As in ( a ), except that the cells were infected with pRetroX-Tight vector encoding VHL-aGFP16. Images were taken using DeltaVision microscopy imaging systems (GE Healthcare) at 60× magnification.
    Pretrox Ptight Tet On Vectors, 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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    99
    Thermo Fisher pcdna3 1
    Degradation of VPS34 through AdPROM results in loss of function. ( a ) GFP-VPS34 knockin HEK293 cells expressing the <t>Tet-transactivator</t> were infected with <t>pRetroX-Tight</t> empty vector control. Following selection, cells were seeded onto glass coverslips and either left untreated or treated with doxycycline (2 µg ml −1 ; 24 h) or VPS34-IN1 (2.5 µM, 1 h). Cells were permealized in liquid nitrogen, and fixed in 3.7% (w/v) paraformaldehyde before staining with fluorescently labelled selective PI3P binding (green) and interaction deficient (red) probes as described in the Material and methods section. Samples were mounted on microscopy slides with mounting media containing DAPI (blue). ( b ) As in ( a ), except that the cells were infected with pRetroX-Tight vector encoding VHL-aGFP16. Images were taken using DeltaVision microscopy imaging systems (GE Healthcare) at 60× magnification.
    Pcdna3 1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 39500 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    TaKaRa pshuttle3 vectors
    Degradation of VPS34 through AdPROM results in loss of function. ( a ) GFP-VPS34 knockin HEK293 cells expressing the <t>Tet-transactivator</t> were infected with <t>pRetroX-Tight</t> empty vector control. Following selection, cells were seeded onto glass coverslips and either left untreated or treated with doxycycline (2 µg ml −1 ; 24 h) or VPS34-IN1 (2.5 µM, 1 h). Cells were permealized in liquid nitrogen, and fixed in 3.7% (w/v) paraformaldehyde before staining with fluorescently labelled selective PI3P binding (green) and interaction deficient (red) probes as described in the Material and methods section. Samples were mounted on microscopy slides with mounting media containing DAPI (blue). ( b ) As in ( a ), except that the cells were infected with pRetroX-Tight vector encoding VHL-aGFP16. Images were taken using DeltaVision microscopy imaging systems (GE Healthcare) at 60× magnification.
    Pshuttle3 Vectors, supplied by TaKaRa, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    TaKaRa pshuttle2
    Degradation of VPS34 through AdPROM results in loss of function. ( a ) GFP-VPS34 knockin HEK293 cells expressing the <t>Tet-transactivator</t> were infected with <t>pRetroX-Tight</t> empty vector control. Following selection, cells were seeded onto glass coverslips and either left untreated or treated with doxycycline (2 µg ml −1 ; 24 h) or VPS34-IN1 (2.5 µM, 1 h). Cells were permealized in liquid nitrogen, and fixed in 3.7% (w/v) paraformaldehyde before staining with fluorescently labelled selective PI3P binding (green) and interaction deficient (red) probes as described in the Material and methods section. Samples were mounted on microscopy slides with mounting media containing DAPI (blue). ( b ) As in ( a ), except that the cells were infected with pRetroX-Tight vector encoding VHL-aGFP16. Images were taken using DeltaVision microscopy imaging systems (GE Healthcare) at 60× magnification.
    Pshuttle2, supplied by TaKaRa, used in various techniques. Bioz Stars score: 86/100, based on 64 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher lipofectamine 2000
    Degradation of VPS34 through AdPROM results in loss of function. ( a ) GFP-VPS34 knockin HEK293 cells expressing the <t>Tet-transactivator</t> were infected with <t>pRetroX-Tight</t> empty vector control. Following selection, cells were seeded onto glass coverslips and either left untreated or treated with doxycycline (2 µg ml −1 ; 24 h) or VPS34-IN1 (2.5 µM, 1 h). Cells were permealized in liquid nitrogen, and fixed in 3.7% (w/v) paraformaldehyde before staining with fluorescently labelled selective PI3P binding (green) and interaction deficient (red) probes as described in the Material and methods section. Samples were mounted on microscopy slides with mounting media containing DAPI (blue). ( b ) As in ( a ), except that the cells were infected with pRetroX-Tight vector encoding VHL-aGFP16. Images were taken using DeltaVision microscopy imaging systems (GE Healthcare) at 60× magnification.
    Lipofectamine 2000, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 377302 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Promega pfn10a act flexi vector
    Degradation of VPS34 through AdPROM results in loss of function. ( a ) GFP-VPS34 knockin HEK293 cells expressing the <t>Tet-transactivator</t> were infected with <t>pRetroX-Tight</t> empty vector control. Following selection, cells were seeded onto glass coverslips and either left untreated or treated with doxycycline (2 µg ml −1 ; 24 h) or VPS34-IN1 (2.5 µM, 1 h). Cells were permealized in liquid nitrogen, and fixed in 3.7% (w/v) paraformaldehyde before staining with fluorescently labelled selective PI3P binding (green) and interaction deficient (red) probes as described in the Material and methods section. Samples were mounted on microscopy slides with mounting media containing DAPI (blue). ( b ) As in ( a ), except that the cells were infected with pRetroX-Tight vector encoding VHL-aGFP16. Images were taken using DeltaVision microscopy imaging systems (GE Healthcare) at 60× magnification.
    Pfn10a Act Flexi Vector, supplied by Promega, used in various techniques. Bioz Stars score: 96/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Promega pgl3 basic vector
    Transcriptional repression by MBD1 isoforms in mammalian cells. (A) Expression of HA-tagged MBD1 isoforms. Four vectors expressing MBD1 isoforms, pCGN-MBD1v1 to pCGN-MBD1v4, were transfected into CHO cells, and a Western blot analysis with an anti-HA epitope monoclonal antibody was performed. (B) Inhibition of p16 , VHL , E-cadherin , and SNRPN gene promoter activities by MBD1 isoforms. Both promoter-inserted <t>pGL3</t> and pCGN-MBD1 vectors in the appropriate combinations were cotransfected into CHO cells. The level of luciferase activity in the cotransfection of unmethylated promoter-inserted pGL3 and pCGN (mock) was normalized to 10,000 in each gene promoter. The average of relative activities from four independent experiments is indicated by each bar. Unmethylated (M−) and methylated (M+) promoter-inserted pGL3 vectors are shown in the upper and lower rows, respectively.
    Pgl3 Basic Vector, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 22990 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    iNtRON Biotechnology muta directtm
    Transcriptional repression by MBD1 isoforms in mammalian cells. (A) Expression of HA-tagged MBD1 isoforms. Four vectors expressing MBD1 isoforms, pCGN-MBD1v1 to pCGN-MBD1v4, were transfected into CHO cells, and a Western blot analysis with an anti-HA epitope monoclonal antibody was performed. (B) Inhibition of p16 , VHL , E-cadherin , and SNRPN gene promoter activities by MBD1 isoforms. Both promoter-inserted <t>pGL3</t> and pCGN-MBD1 vectors in the appropriate combinations were cotransfected into CHO cells. The level of luciferase activity in the cotransfection of unmethylated promoter-inserted pGL3 and pCGN (mock) was normalized to 10,000 in each gene promoter. The average of relative activities from four independent experiments is indicated by each bar. Unmethylated (M−) and methylated (M+) promoter-inserted pGL3 vectors are shown in the upper and lower rows, respectively.
    Muta Directtm, supplied by iNtRON Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    TaKaRa y187 yeast strain
    Transcriptional repression by MBD1 isoforms in mammalian cells. (A) Expression of HA-tagged MBD1 isoforms. Four vectors expressing MBD1 isoforms, pCGN-MBD1v1 to pCGN-MBD1v4, were transfected into CHO cells, and a Western blot analysis with an anti-HA epitope monoclonal antibody was performed. (B) Inhibition of p16 , VHL , E-cadherin , and SNRPN gene promoter activities by MBD1 isoforms. Both promoter-inserted <t>pGL3</t> and pCGN-MBD1 vectors in the appropriate combinations were cotransfected into CHO cells. The level of luciferase activity in the cotransfection of unmethylated promoter-inserted pGL3 and pCGN (mock) was normalized to 10,000 in each gene promoter. The average of relative activities from four independent experiments is indicated by each bar. Unmethylated (M−) and methylated (M+) promoter-inserted pGL3 vectors are shown in the upper and lower rows, respectively.
    Y187 Yeast Strain, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 260 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Promega pgl3 control vector
    Transcriptional repression by MBD1 isoforms in mammalian cells. (A) Expression of HA-tagged MBD1 isoforms. Four vectors expressing MBD1 isoforms, pCGN-MBD1v1 to pCGN-MBD1v4, were transfected into CHO cells, and a Western blot analysis with an anti-HA epitope monoclonal antibody was performed. (B) Inhibition of p16 , VHL , E-cadherin , and SNRPN gene promoter activities by MBD1 isoforms. Both promoter-inserted <t>pGL3</t> and pCGN-MBD1 vectors in the appropriate combinations were cotransfected into CHO cells. The level of luciferase activity in the cotransfection of unmethylated promoter-inserted pGL3 and pCGN (mock) was normalized to 10,000 in each gene promoter. The average of relative activities from four independent experiments is indicated by each bar. Unmethylated (M−) and methylated (M+) promoter-inserted pGL3 vectors are shown in the upper and lower rows, respectively.
    Pgl3 Control Vector, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 6315 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher psuper vector
    Transcriptional repression by MBD1 isoforms in mammalian cells. (A) Expression of HA-tagged MBD1 isoforms. Four vectors expressing MBD1 isoforms, pCGN-MBD1v1 to pCGN-MBD1v4, were transfected into CHO cells, and a Western blot analysis with an anti-HA epitope monoclonal antibody was performed. (B) Inhibition of p16 , VHL , E-cadherin , and SNRPN gene promoter activities by MBD1 isoforms. Both promoter-inserted <t>pGL3</t> and pCGN-MBD1 vectors in the appropriate combinations were cotransfected into CHO cells. The level of luciferase activity in the cotransfection of unmethylated promoter-inserted pGL3 and pCGN (mock) was normalized to 10,000 in each gene promoter. The average of relative activities from four independent experiments is indicated by each bar. Unmethylated (M−) and methylated (M+) promoter-inserted pGL3 vectors are shown in the upper and lower rows, respectively.
    Psuper Vector, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 194 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    TaKaRa yeast two hybrid screen
    Transcriptional repression by MBD1 isoforms in mammalian cells. (A) Expression of HA-tagged MBD1 isoforms. Four vectors expressing MBD1 isoforms, pCGN-MBD1v1 to pCGN-MBD1v4, were transfected into CHO cells, and a Western blot analysis with an anti-HA epitope monoclonal antibody was performed. (B) Inhibition of p16 , VHL , E-cadherin , and SNRPN gene promoter activities by MBD1 isoforms. Both promoter-inserted <t>pGL3</t> and pCGN-MBD1 vectors in the appropriate combinations were cotransfected into CHO cells. The level of luciferase activity in the cotransfection of unmethylated promoter-inserted pGL3 and pCGN (mock) was normalized to 10,000 in each gene promoter. The average of relative activities from four independent experiments is indicated by each bar. Unmethylated (M−) and methylated (M+) promoter-inserted pGL3 vectors are shown in the upper and lower rows, respectively.
    Yeast Two Hybrid Screen, supplied by TaKaRa, used in various techniques. Bioz Stars score: 96/100, based on 1518 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Millipore pflag cmv1
    Transcriptional repression by MBD1 isoforms in mammalian cells. (A) Expression of HA-tagged MBD1 isoforms. Four vectors expressing MBD1 isoforms, pCGN-MBD1v1 to pCGN-MBD1v4, were transfected into CHO cells, and a Western blot analysis with an anti-HA epitope monoclonal antibody was performed. (B) Inhibition of p16 , VHL , E-cadherin , and SNRPN gene promoter activities by MBD1 isoforms. Both promoter-inserted <t>pGL3</t> and pCGN-MBD1 vectors in the appropriate combinations were cotransfected into CHO cells. The level of luciferase activity in the cotransfection of unmethylated promoter-inserted pGL3 and pCGN (mock) was normalized to 10,000 in each gene promoter. The average of relative activities from four independent experiments is indicated by each bar. Unmethylated (M−) and methylated (M+) promoter-inserted pGL3 vectors are shown in the upper and lower rows, respectively.
    Pflag Cmv1, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 117 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    TaKaRa lexa dna binding domain
    Transcriptional repression by MBD1 isoforms in mammalian cells. (A) Expression of HA-tagged MBD1 isoforms. Four vectors expressing MBD1 isoforms, pCGN-MBD1v1 to pCGN-MBD1v4, were transfected into CHO cells, and a Western blot analysis with an anti-HA epitope monoclonal antibody was performed. (B) Inhibition of p16 , VHL , E-cadherin , and SNRPN gene promoter activities by MBD1 isoforms. Both promoter-inserted <t>pGL3</t> and pCGN-MBD1 vectors in the appropriate combinations were cotransfected into CHO cells. The level of luciferase activity in the cotransfection of unmethylated promoter-inserted pGL3 and pCGN (mock) was normalized to 10,000 in each gene promoter. The average of relative activities from four independent experiments is indicated by each bar. Unmethylated (M−) and methylated (M+) promoter-inserted pGL3 vectors are shown in the upper and lower rows, respectively.
    Lexa Dna Binding Domain, supplied by TaKaRa, used in various techniques. Bioz Stars score: 89/100, based on 104 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    TaKaRa yeast two hybrid system 3
    Transcriptional repression by MBD1 isoforms in mammalian cells. (A) Expression of HA-tagged MBD1 isoforms. Four vectors expressing MBD1 isoforms, pCGN-MBD1v1 to pCGN-MBD1v4, were transfected into CHO cells, and a Western blot analysis with an anti-HA epitope monoclonal antibody was performed. (B) Inhibition of p16 , VHL , E-cadherin , and SNRPN gene promoter activities by MBD1 isoforms. Both promoter-inserted <t>pGL3</t> and pCGN-MBD1 vectors in the appropriate combinations were cotransfected into CHO cells. The level of luciferase activity in the cotransfection of unmethylated promoter-inserted pGL3 and pCGN (mock) was normalized to 10,000 in each gene promoter. The average of relative activities from four independent experiments is indicated by each bar. Unmethylated (M−) and methylated (M+) promoter-inserted pGL3 vectors are shown in the upper and lower rows, respectively.
    Yeast Two Hybrid System 3, supplied by TaKaRa, used in various techniques. Bioz Stars score: 86/100, based on 61 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Stratagene pcmv5 mammalian expression vector
    Transcriptional repression by MBD1 isoforms in mammalian cells. (A) Expression of HA-tagged MBD1 isoforms. Four vectors expressing MBD1 isoforms, pCGN-MBD1v1 to pCGN-MBD1v4, were transfected into CHO cells, and a Western blot analysis with an anti-HA epitope monoclonal antibody was performed. (B) Inhibition of p16 , VHL , E-cadherin , and SNRPN gene promoter activities by MBD1 isoforms. Both promoter-inserted <t>pGL3</t> and pCGN-MBD1 vectors in the appropriate combinations were cotransfected into CHO cells. The level of luciferase activity in the cotransfection of unmethylated promoter-inserted pGL3 and pCGN (mock) was normalized to 10,000 in each gene promoter. The average of relative activities from four independent experiments is indicated by each bar. Unmethylated (M−) and methylated (M+) promoter-inserted pGL3 vectors are shown in the upper and lower rows, respectively.
    Pcmv5 Mammalian Expression Vector, supplied by Stratagene, used in various techniques. Bioz Stars score: 86/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene quickchange ii kit
    Transcriptional repression by MBD1 isoforms in mammalian cells. (A) Expression of HA-tagged MBD1 isoforms. Four vectors expressing MBD1 isoforms, pCGN-MBD1v1 to pCGN-MBD1v4, were transfected into CHO cells, and a Western blot analysis with an anti-HA epitope monoclonal antibody was performed. (B) Inhibition of p16 , VHL , E-cadherin , and SNRPN gene promoter activities by MBD1 isoforms. Both promoter-inserted <t>pGL3</t> and pCGN-MBD1 vectors in the appropriate combinations were cotransfected into CHO cells. The level of luciferase activity in the cotransfection of unmethylated promoter-inserted pGL3 and pCGN (mock) was normalized to 10,000 in each gene promoter. The average of relative activities from four independent experiments is indicated by each bar. Unmethylated (M−) and methylated (M+) promoter-inserted pGL3 vectors are shown in the upper and lower rows, respectively.
    Quickchange Ii Kit, supplied by Stratagene, used in various techniques. Bioz Stars score: 97/100, based on 428 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC sw839 vhl cell lines
    Transcriptional repression by MBD1 isoforms in mammalian cells. (A) Expression of HA-tagged MBD1 isoforms. Four vectors expressing MBD1 isoforms, pCGN-MBD1v1 to pCGN-MBD1v4, were transfected into CHO cells, and a Western blot analysis with an anti-HA epitope monoclonal antibody was performed. (B) Inhibition of p16 , VHL , E-cadherin , and SNRPN gene promoter activities by MBD1 isoforms. Both promoter-inserted <t>pGL3</t> and pCGN-MBD1 vectors in the appropriate combinations were cotransfected into CHO cells. The level of luciferase activity in the cotransfection of unmethylated promoter-inserted pGL3 and pCGN (mock) was normalized to 10,000 in each gene promoter. The average of relative activities from four independent experiments is indicated by each bar. Unmethylated (M−) and methylated (M+) promoter-inserted pGL3 vectors are shown in the upper and lower rows, respectively.
    Sw839 Vhl Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen rneasy plus mini kit qiagen valencia ca
    Transcriptional repression by MBD1 isoforms in mammalian cells. (A) Expression of HA-tagged MBD1 isoforms. Four vectors expressing MBD1 isoforms, pCGN-MBD1v1 to pCGN-MBD1v4, were transfected into CHO cells, and a Western blot analysis with an anti-HA epitope monoclonal antibody was performed. (B) Inhibition of p16 , VHL , E-cadherin , and SNRPN gene promoter activities by MBD1 isoforms. Both promoter-inserted <t>pGL3</t> and pCGN-MBD1 vectors in the appropriate combinations were cotransfected into CHO cells. The level of luciferase activity in the cotransfection of unmethylated promoter-inserted pGL3 and pCGN (mock) was normalized to 10,000 in each gene promoter. The average of relative activities from four independent experiments is indicated by each bar. Unmethylated (M−) and methylated (M+) promoter-inserted pGL3 vectors are shown in the upper and lower rows, respectively.
    Rneasy Plus Mini Kit Qiagen Valencia Ca, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    EGFR -activating mutations are associated with elevated MET and HIF-1α levels in NSCLC cell lines. ( a ) Gene expression analysis was performed on gene arrays of 53 NSCLC lines. MET expression was elevated in NSCLC cell lines harboring EGFR -activating mutations; * P = 0.002. ( b ) MET expression in 53 NSCLC cell lines with high EGFR gene copy number ( > 4 copies) vs low copy number (

    Journal: Oncogene

    Article Title: Epidermal growth factor receptor regulates MET levels and invasiveness through hypoxia-inducible factor-1? in non-small cell lung cancer cells

    doi: 10.1038/onc.2010.16

    Figure Lengend Snippet: EGFR -activating mutations are associated with elevated MET and HIF-1α levels in NSCLC cell lines. ( a ) Gene expression analysis was performed on gene arrays of 53 NSCLC lines. MET expression was elevated in NSCLC cell lines harboring EGFR -activating mutations; * P = 0.002. ( b ) MET expression in 53 NSCLC cell lines with high EGFR gene copy number ( > 4 copies) vs low copy number (

    Article Snippet: HIF-1α cDNA (OriGene Technologies Inc., Rockville, MD, USA) was subcloned into the pcDNA3.1 vector with a flag tagged in the N-terminal, and the HIF-1α mutant with proline to alanine substitutions positions 402 and 564 (HA- HIF-1α P402A;P564A), which are known VHL-binding sites, was constructed as described ( ).

    Techniques: Expressing, Low Copy Number

    Degradation of VPS34 through AdPROM results in loss of function. ( a ) GFP-VPS34 knockin HEK293 cells expressing the Tet-transactivator were infected with pRetroX-Tight empty vector control. Following selection, cells were seeded onto glass coverslips and either left untreated or treated with doxycycline (2 µg ml −1 ; 24 h) or VPS34-IN1 (2.5 µM, 1 h). Cells were permealized in liquid nitrogen, and fixed in 3.7% (w/v) paraformaldehyde before staining with fluorescently labelled selective PI3P binding (green) and interaction deficient (red) probes as described in the Material and methods section. Samples were mounted on microscopy slides with mounting media containing DAPI (blue). ( b ) As in ( a ), except that the cells were infected with pRetroX-Tight vector encoding VHL-aGFP16. Images were taken using DeltaVision microscopy imaging systems (GE Healthcare) at 60× magnification.

    Journal: Open Biology

    Article Title: An affinity-directed protein missile system for targeted proteolysis

    doi: 10.1098/rsob.160255

    Figure Lengend Snippet: Degradation of VPS34 through AdPROM results in loss of function. ( a ) GFP-VPS34 knockin HEK293 cells expressing the Tet-transactivator were infected with pRetroX-Tight empty vector control. Following selection, cells were seeded onto glass coverslips and either left untreated or treated with doxycycline (2 µg ml −1 ; 24 h) or VPS34-IN1 (2.5 µM, 1 h). Cells were permealized in liquid nitrogen, and fixed in 3.7% (w/v) paraformaldehyde before staining with fluorescently labelled selective PI3P binding (green) and interaction deficient (red) probes as described in the Material and methods section. Samples were mounted on microscopy slides with mounting media containing DAPI (blue). ( b ) As in ( a ), except that the cells were infected with pRetroX-Tight vector encoding VHL-aGFP16. Images were taken using DeltaVision microscopy imaging systems (GE Healthcare) at 60× magnification.

    Article Snippet: The cDNAs encoding GFP (DU32961), anti-GFP nanobodies (aGFP-DU54218; aGFP16-DU54238) [ , ], human VHL (DU54023), aGFP-VHL (DU54023), VHL-aGFP (DU54221) and VHL-aGFP16 (DU54294) were cloned into pBABED-Puro vectors (Cell Biolabs, modified) for constitutive expression and pRetroX-pTight Tet-ON vectors (Clontech) for tetracycline-inducible expression.

    Techniques: Knock-In, Expressing, Infection, Plasmid Preparation, Selection, Staining, Binding Assay, Microscopy, Imaging

    Adapting AdPROM for tetracycline-inducible degradation of target proteins: GFP-VPS34 knockin HEK293 cells were first infected with the pRetroX-Tet-ON advanced vector (Clontech) and selected for the expression of Tet-transactivator. Cells were then infected with either pRetroX-Tight empty vector control or pRetroX-Tight vector encoding VHL-aGFP16. Cells were then treated with 2 µg ml −1 doxycycline for the indicated time points prior to lysis. Extracts (20 µg protein) were resolved by SDS–PAGE, transferred to PVDF membranes and subjected to western blotting, using antibodies against GFP, VPS34, UVRAG and VHL, as indicated. Anti-GAPDH antibody was included as a loading control.

    Journal: Open Biology

    Article Title: An affinity-directed protein missile system for targeted proteolysis

    doi: 10.1098/rsob.160255

    Figure Lengend Snippet: Adapting AdPROM for tetracycline-inducible degradation of target proteins: GFP-VPS34 knockin HEK293 cells were first infected with the pRetroX-Tet-ON advanced vector (Clontech) and selected for the expression of Tet-transactivator. Cells were then infected with either pRetroX-Tight empty vector control or pRetroX-Tight vector encoding VHL-aGFP16. Cells were then treated with 2 µg ml −1 doxycycline for the indicated time points prior to lysis. Extracts (20 µg protein) were resolved by SDS–PAGE, transferred to PVDF membranes and subjected to western blotting, using antibodies against GFP, VPS34, UVRAG and VHL, as indicated. Anti-GAPDH antibody was included as a loading control.

    Article Snippet: The cDNAs encoding GFP (DU32961), anti-GFP nanobodies (aGFP-DU54218; aGFP16-DU54238) [ , ], human VHL (DU54023), aGFP-VHL (DU54023), VHL-aGFP (DU54221) and VHL-aGFP16 (DU54294) were cloned into pBABED-Puro vectors (Cell Biolabs, modified) for constitutive expression and pRetroX-pTight Tet-ON vectors (Clontech) for tetracycline-inducible expression.

    Techniques: Knock-In, Infection, Plasmid Preparation, Expressing, Lysis, SDS Page, Western Blot

    The affinity-directed protein missile (AdPROM) system degrades target proteins in different cell lines. ( a ) Schematic describes how the CUL2-E3 ligase complex results in ubiquitylation and degradation of its native target, hydroxy-proline modified HIF1α, under normoxic conditions. ( b ) Schematic of the exploitation of the CUL2 E3 ligase machinery for AdPROM using anti-GFP nanobody (aGFP) to degrade GFP-tagged proteins. ( c ) Strategy for rapidly knocking in a GFP tag on target proteins in somatic cells using CRISPR/Cas9. ( d ) Western blot analysis of extracts from PAWS1-GFP and GFP-VPS34 knockin U2OS and HEK293 cells, respectively, using the indicated antibodies. ( e ) Schematic shows the application of AdPROM using pBABED-Puro (for constitutive expression) or pRetroX-TetON (for Tet-inducible expression) retroviral infection systems to introduce the VHL-aGFP in GFP-knockin cells. ( f ) A proof-of-principle demonstration of the efficacy of the AdPROM system in the degradation of GFP-VPS34 and PAWS1-GFP from knockin HEK293 and U2OS cells, respectively. Cells infected with control retroviruses (GFP) or retroviruses encoding aGFP-VHL or VHL-aGFP were lysed. Extracts (20 µg protein) were subjected to resolution by SDS–PAGE and transferred to PVDF membranes, which were analysed by western blotting with the indicated antibodies.

    Journal: Open Biology

    Article Title: An affinity-directed protein missile system for targeted proteolysis

    doi: 10.1098/rsob.160255

    Figure Lengend Snippet: The affinity-directed protein missile (AdPROM) system degrades target proteins in different cell lines. ( a ) Schematic describes how the CUL2-E3 ligase complex results in ubiquitylation and degradation of its native target, hydroxy-proline modified HIF1α, under normoxic conditions. ( b ) Schematic of the exploitation of the CUL2 E3 ligase machinery for AdPROM using anti-GFP nanobody (aGFP) to degrade GFP-tagged proteins. ( c ) Strategy for rapidly knocking in a GFP tag on target proteins in somatic cells using CRISPR/Cas9. ( d ) Western blot analysis of extracts from PAWS1-GFP and GFP-VPS34 knockin U2OS and HEK293 cells, respectively, using the indicated antibodies. ( e ) Schematic shows the application of AdPROM using pBABED-Puro (for constitutive expression) or pRetroX-TetON (for Tet-inducible expression) retroviral infection systems to introduce the VHL-aGFP in GFP-knockin cells. ( f ) A proof-of-principle demonstration of the efficacy of the AdPROM system in the degradation of GFP-VPS34 and PAWS1-GFP from knockin HEK293 and U2OS cells, respectively. Cells infected with control retroviruses (GFP) or retroviruses encoding aGFP-VHL or VHL-aGFP were lysed. Extracts (20 µg protein) were subjected to resolution by SDS–PAGE and transferred to PVDF membranes, which were analysed by western blotting with the indicated antibodies.

    Article Snippet: The cDNAs encoding GFP (DU32961), anti-GFP nanobodies (aGFP-DU54218; aGFP16-DU54238) [ , ], human VHL (DU54023), aGFP-VHL (DU54023), VHL-aGFP (DU54221) and VHL-aGFP16 (DU54294) were cloned into pBABED-Puro vectors (Cell Biolabs, modified) for constitutive expression and pRetroX-pTight Tet-ON vectors (Clontech) for tetracycline-inducible expression.

    Techniques: Modification, CRISPR, Western Blot, Knock-In, Expressing, Infection, Introduce, SDS Page

    Transcriptional repression by MBD1 isoforms in mammalian cells. (A) Expression of HA-tagged MBD1 isoforms. Four vectors expressing MBD1 isoforms, pCGN-MBD1v1 to pCGN-MBD1v4, were transfected into CHO cells, and a Western blot analysis with an anti-HA epitope monoclonal antibody was performed. (B) Inhibition of p16 , VHL , E-cadherin , and SNRPN gene promoter activities by MBD1 isoforms. Both promoter-inserted pGL3 and pCGN-MBD1 vectors in the appropriate combinations were cotransfected into CHO cells. The level of luciferase activity in the cotransfection of unmethylated promoter-inserted pGL3 and pCGN (mock) was normalized to 10,000 in each gene promoter. The average of relative activities from four independent experiments is indicated by each bar. Unmethylated (M−) and methylated (M+) promoter-inserted pGL3 vectors are shown in the upper and lower rows, respectively.

    Journal: Molecular and Cellular Biology

    Article Title: Methylation-Mediated Transcriptional Silencing in Euchromatin by Methyl-CpG Binding Protein MBD1 Isoforms

    doi:

    Figure Lengend Snippet: Transcriptional repression by MBD1 isoforms in mammalian cells. (A) Expression of HA-tagged MBD1 isoforms. Four vectors expressing MBD1 isoforms, pCGN-MBD1v1 to pCGN-MBD1v4, were transfected into CHO cells, and a Western blot analysis with an anti-HA epitope monoclonal antibody was performed. (B) Inhibition of p16 , VHL , E-cadherin , and SNRPN gene promoter activities by MBD1 isoforms. Both promoter-inserted pGL3 and pCGN-MBD1 vectors in the appropriate combinations were cotransfected into CHO cells. The level of luciferase activity in the cotransfection of unmethylated promoter-inserted pGL3 and pCGN (mock) was normalized to 10,000 in each gene promoter. The average of relative activities from four independent experiments is indicated by each bar. Unmethylated (M−) and methylated (M+) promoter-inserted pGL3 vectors are shown in the upper and lower rows, respectively.

    Article Snippet: The PCR products amplified from the promoter regions of p16 , E-cadherin , VHL , and SNRPN genes were double digested with Nhe I and Hin dIII and ligated upstream of a luciferase cDNA in a pGL3-Basic vector (Promega, Madison, Wis.).

    Techniques: Expressing, Transfection, Western Blot, Inhibition, Luciferase, Activity Assay, Cotransfection, Methylation

    Transcriptional regulation by MBD1 isoforms in Drosophila SL2 cells. (A) Unmethylated (M−) or methylated (M+) pGL3 construct (pGL3-p16 and pGL3-SNRPN or pGL3-VHL and pGL3-SNRPN) was cotransfected in combination with mock A5C vector (solid bars), pPacSp1 expressing transcription factor Sp1 (hatched bars on the left), or pAc5.1-E2F1 expressing transcription factor E2F1 (hatched bars on the right) into SL2 cells. The level of luciferase activity in the presence of unmethylated pGL3 and mock vectors was normalized to 1 for each gene promoter. (B) Methylated (solid bars) or unmethylated (hatched bars) promoter-inserted pGL3 vector was cotransfected with pPacSp1 and MBD1-expressing plasmids (pAc5.1-MBD1v1 to pAc5.1-MBD1v4) or insertless plasmid (mock). The luciferase activity of unmethylated pGL3 in the combination of pPacSp1 and mock was normalized to 100 for each gene promoter, and the relative luciferase activities are presented.

    Journal: Molecular and Cellular Biology

    Article Title: Methylation-Mediated Transcriptional Silencing in Euchromatin by Methyl-CpG Binding Protein MBD1 Isoforms

    doi:

    Figure Lengend Snippet: Transcriptional regulation by MBD1 isoforms in Drosophila SL2 cells. (A) Unmethylated (M−) or methylated (M+) pGL3 construct (pGL3-p16 and pGL3-SNRPN or pGL3-VHL and pGL3-SNRPN) was cotransfected in combination with mock A5C vector (solid bars), pPacSp1 expressing transcription factor Sp1 (hatched bars on the left), or pAc5.1-E2F1 expressing transcription factor E2F1 (hatched bars on the right) into SL2 cells. The level of luciferase activity in the presence of unmethylated pGL3 and mock vectors was normalized to 1 for each gene promoter. (B) Methylated (solid bars) or unmethylated (hatched bars) promoter-inserted pGL3 vector was cotransfected with pPacSp1 and MBD1-expressing plasmids (pAc5.1-MBD1v1 to pAc5.1-MBD1v4) or insertless plasmid (mock). The luciferase activity of unmethylated pGL3 in the combination of pPacSp1 and mock was normalized to 100 for each gene promoter, and the relative luciferase activities are presented.

    Article Snippet: The PCR products amplified from the promoter regions of p16 , E-cadherin , VHL , and SNRPN genes were double digested with Nhe I and Hin dIII and ligated upstream of a luciferase cDNA in a pGL3-Basic vector (Promega, Madison, Wis.).

    Techniques: Methylation, Construct, Plasmid Preparation, Expressing, Luciferase, Activity Assay

    Effect of MBD1 on promoter-associated CpG islands of p16 , VHL , E-cadherin , and SNRPN genes. (A) PCR-amplified DNA fragments from the gene promoters were subcloned upstream of a luciferase cDNA in a pGL3-Basic vector. The position of the transcription start site and the oligonucleotide primers DF and ER to amplify DNA fragments utilized for the in vitro transcription assay are shown by solid and open arrows, respectively. The presence of CpG dinucleotides within the inserted promoter sequences of four genes is indicated by vertical lines. (B) Band shift of methylated DNAs complexed with MBD1. Unmethylated (M−) and methylated (M+) DNA fragments containing CpG islands from these genes were incubated with GST-fused MBD1v1 or GST alone. The in vitro methylation of CpG sequences was performed with Sss I methyltransferase. +, present; −, absent. (C) Transcriptional repression by MBD1 in a methylation-dependent manner. GST-MBD1 was incubated with either unmethylated (M−) or methylated (M+) DNA fragments which were PCR amplified from the promoter-inserted pGL3 vectors with primers DF and ER. Transcripts from the promoters were synthesized in HeLa nuclear extract and detected by the primer extension method with a radiolabeled ER primer. The amount of a predicted cDNA product was measured by a Bioimaging analyzer, MacBAS version 2.51, and the relative amount of transcript compared with that of the unmethylated promoter without GST-MBD1 is indicated below each panel.

    Journal: Molecular and Cellular Biology

    Article Title: Methylation-Mediated Transcriptional Silencing in Euchromatin by Methyl-CpG Binding Protein MBD1 Isoforms

    doi:

    Figure Lengend Snippet: Effect of MBD1 on promoter-associated CpG islands of p16 , VHL , E-cadherin , and SNRPN genes. (A) PCR-amplified DNA fragments from the gene promoters were subcloned upstream of a luciferase cDNA in a pGL3-Basic vector. The position of the transcription start site and the oligonucleotide primers DF and ER to amplify DNA fragments utilized for the in vitro transcription assay are shown by solid and open arrows, respectively. The presence of CpG dinucleotides within the inserted promoter sequences of four genes is indicated by vertical lines. (B) Band shift of methylated DNAs complexed with MBD1. Unmethylated (M−) and methylated (M+) DNA fragments containing CpG islands from these genes were incubated with GST-fused MBD1v1 or GST alone. The in vitro methylation of CpG sequences was performed with Sss I methyltransferase. +, present; −, absent. (C) Transcriptional repression by MBD1 in a methylation-dependent manner. GST-MBD1 was incubated with either unmethylated (M−) or methylated (M+) DNA fragments which were PCR amplified from the promoter-inserted pGL3 vectors with primers DF and ER. Transcripts from the promoters were synthesized in HeLa nuclear extract and detected by the primer extension method with a radiolabeled ER primer. The amount of a predicted cDNA product was measured by a Bioimaging analyzer, MacBAS version 2.51, and the relative amount of transcript compared with that of the unmethylated promoter without GST-MBD1 is indicated below each panel.

    Article Snippet: The PCR products amplified from the promoter regions of p16 , E-cadherin , VHL , and SNRPN genes were double digested with Nhe I and Hin dIII and ligated upstream of a luciferase cDNA in a pGL3-Basic vector (Promega, Madison, Wis.).

    Techniques: Polymerase Chain Reaction, Amplification, Luciferase, Plasmid Preparation, In Vitro, Electrophoretic Mobility Shift Assay, Methylation, Incubation, Synthesized