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destination luciferase reporter vector pgl4 23  (Addgene inc)


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    Addgene inc destination luciferase reporter vector pgl4 23
    Destination Luciferase Reporter Vector Pgl4 23, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/reporter+vector/pGL4%2E23-GW+(Plasmid+%2360323)/pm41871099-769-27-34
    Average 93 stars, based on 19 article reviews
    destination luciferase reporter vector pgl4 23 - by Bioz Stars, 2026-09
    93/100 stars

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    Related Articles

    Plasmid Preparation:

    Article Title: Polypeptide containing DNA-binding domain
    Article Snippet: .. First, a reporter vector was prepared in which a target sequence of TALEN of interest was inserted into a reporter vector (pGL4-SSA; Addgene) wherein a segmented firefly luciferase gene was linked downstream CMV promoter. .. The target sequence of TALEN was prepared by annealing synthetic oligonucleotides and was inserted into pGL4-SSA vector treated with BsaI using Ligation-Convenience Kit (NIPPON GENE CO., LTD.).

    Article Title: Massively Parallel Reporter Assays for High-Throughput In Vivo Analysis of Cis-Regulatory Elements.
    Article Snippet: The first, our work with genomic regions bound by multiple core cardiac transcription factors [42], featured a library of 2700 regions, each 400 bp in length, generated by annealing overlapping oligos. .. Regions were cloned into the 3′ UTR of a scAAV Mlc2v promoter-containing reporter vector (Addgene #182649), which was packaged into the AAV9 capsid and delivered to newborn mice. ..

    Sequencing:

    Article Title: Polypeptide containing DNA-binding domain
    Article Snippet: .. First, a reporter vector was prepared in which a target sequence of TALEN of interest was inserted into a reporter vector (pGL4-SSA; Addgene) wherein a segmented firefly luciferase gene was linked downstream CMV promoter. .. The target sequence of TALEN was prepared by annealing synthetic oligonucleotides and was inserted into pGL4-SSA vector treated with BsaI using Ligation-Convenience Kit (NIPPON GENE CO., LTD.).

    Article Title: Chemical Specification of E3 Ubiquitin Ligase Engagement by Cysteine-Reactive Chemistry
    Article Snippet: A mixture containing 1:100 NanoGlo substrate:FluoroBrite DMEM was then added to each well and the plate was immediately read on a PHERAStar plate reader (BMG LABTECH) using a LUM 610-LP 450-80 optical module. .. BRD4BD1 and BRD4BD2 reporter cell line generation Reporter constructs were generated by BsmBI (New England Biolabs) digestion of Cilantro (PGK.BsmBICloneSite.FlexibleLinker.eGFP.IRES.mCherry.cppt.EF1α.PuroR, Addgene 74450) reporter vector and the insert containing BRD4BD1 or BRD4BD2 coding sequence, followed by ligation with T4 DNA Ligase (New England Biolabs). .. Constructs were transformed into Stbl3 E. coli and purified using the MiniPrep Kit (Qiagen), and sequences were confirmed by Sanger sequencing (Quintara Biosciences Service).

    Luciferase:

    Article Title: Polypeptide containing DNA-binding domain
    Article Snippet: .. First, a reporter vector was prepared in which a target sequence of TALEN of interest was inserted into a reporter vector (pGL4-SSA; Addgene) wherein a segmented firefly luciferase gene was linked downstream CMV promoter. .. The target sequence of TALEN was prepared by annealing synthetic oligonucleotides and was inserted into pGL4-SSA vector treated with BsaI using Ligation-Convenience Kit (NIPPON GENE CO., LTD.).

    Construct:

    Article Title: Chemical Specification of E3 Ubiquitin Ligase Engagement by Cysteine-Reactive Chemistry
    Article Snippet: A mixture containing 1:100 NanoGlo substrate:FluoroBrite DMEM was then added to each well and the plate was immediately read on a PHERAStar plate reader (BMG LABTECH) using a LUM 610-LP 450-80 optical module. .. BRD4BD1 and BRD4BD2 reporter cell line generation Reporter constructs were generated by BsmBI (New England Biolabs) digestion of Cilantro (PGK.BsmBICloneSite.FlexibleLinker.eGFP.IRES.mCherry.cppt.EF1α.PuroR, Addgene 74450) reporter vector and the insert containing BRD4BD1 or BRD4BD2 coding sequence, followed by ligation with T4 DNA Ligase (New England Biolabs). .. Constructs were transformed into Stbl3 E. coli and purified using the MiniPrep Kit (Qiagen), and sequences were confirmed by Sanger sequencing (Quintara Biosciences Service).

    Generated:

    Article Title: Chemical Specification of E3 Ubiquitin Ligase Engagement by Cysteine-Reactive Chemistry
    Article Snippet: A mixture containing 1:100 NanoGlo substrate:FluoroBrite DMEM was then added to each well and the plate was immediately read on a PHERAStar plate reader (BMG LABTECH) using a LUM 610-LP 450-80 optical module. .. BRD4BD1 and BRD4BD2 reporter cell line generation Reporter constructs were generated by BsmBI (New England Biolabs) digestion of Cilantro (PGK.BsmBICloneSite.FlexibleLinker.eGFP.IRES.mCherry.cppt.EF1α.PuroR, Addgene 74450) reporter vector and the insert containing BRD4BD1 or BRD4BD2 coding sequence, followed by ligation with T4 DNA Ligase (New England Biolabs). .. Constructs were transformed into Stbl3 E. coli and purified using the MiniPrep Kit (Qiagen), and sequences were confirmed by Sanger sequencing (Quintara Biosciences Service).

    Ligation:

    Article Title: Chemical Specification of E3 Ubiquitin Ligase Engagement by Cysteine-Reactive Chemistry
    Article Snippet: A mixture containing 1:100 NanoGlo substrate:FluoroBrite DMEM was then added to each well and the plate was immediately read on a PHERAStar plate reader (BMG LABTECH) using a LUM 610-LP 450-80 optical module. .. BRD4BD1 and BRD4BD2 reporter cell line generation Reporter constructs were generated by BsmBI (New England Biolabs) digestion of Cilantro (PGK.BsmBICloneSite.FlexibleLinker.eGFP.IRES.mCherry.cppt.EF1α.PuroR, Addgene 74450) reporter vector and the insert containing BRD4BD1 or BRD4BD2 coding sequence, followed by ligation with T4 DNA Ligase (New England Biolabs). .. Constructs were transformed into Stbl3 E. coli and purified using the MiniPrep Kit (Qiagen), and sequences were confirmed by Sanger sequencing (Quintara Biosciences Service).

    Clone Assay:

    Article Title: Massively Parallel Reporter Assays for High-Throughput In Vivo Analysis of Cis-Regulatory Elements.
    Article Snippet: The first, our work with genomic regions bound by multiple core cardiac transcription factors [42], featured a library of 2700 regions, each 400 bp in length, generated by annealing overlapping oligos. .. Regions were cloned into the 3′ UTR of a scAAV Mlc2v promoter-containing reporter vector (Addgene #182649), which was packaged into the AAV9 capsid and delivered to newborn mice. ..



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    Detachment‐induced NNMT is upregulated by the FAK‐STAT3 axis. (A) Representative image of the positive correlation between FAK and STAT3 in breast cancer and pan‐cancer according to the GEPIA 2 database analysis. (B) Representative results of STAT3, P‐STAT3, FAK, and P‐FAK proteins by WB in the detached BT‐549, MCF‐7, and MDA‐MB‐231 cells. (C,D) The protein and mRNA levels of NNMT and P‐STAT3 after treating detached cells with FAK inhibitor PF‐562271 (10 µ m ) and P‐STAT3 inhibitor C188‐9 (10 µ m ) for 48 h by WB and QPCR. ** p < 0.01, *** p < 0.001, **** p < 0.0001. (E,F) The protein and mRNA levels of NNMT and P‐STAT3 after treating detached cells with FAK inhibitor PF‐562271 (10 µ m for 48 h) and P‐STAT3 activator (1 µ m for 24 h) by WB and QPCR. ** p < 0.01, *** p < 0.001, **** p < 0.0001. (G,H) ChIP assay using P‐STAT3 antibodies in MDA‐MB‐231 cells cultured under suspension conditions was performed, followed by PCR and qPCR, which showed P‐STAT3 enrichment at the NNMT promoter vs. IgG control. ** p < 0.01. I: <t>Dual‐luciferase</t> reporter assays showed that co‐transfection with the full‐length NNMT promoter and a STAT3‐expressing plasmid significantly enhanced luciferase activity, whereas this enhancement was abolished when a truncated NNMT promoter was used. Data were presented as mean ± SEM. **** p < 0.0001.
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    Detachment‐induced NNMT is upregulated by the FAK‐STAT3 axis. (A) Representative image of the positive correlation between FAK and STAT3 in breast cancer and pan‐cancer according to the GEPIA 2 database analysis. (B) Representative results of STAT3, P‐STAT3, FAK, and P‐FAK proteins by WB in the detached BT‐549, MCF‐7, and MDA‐MB‐231 cells. (C,D) The protein and mRNA levels of NNMT and P‐STAT3 after treating detached cells with FAK inhibitor PF‐562271 (10 µ m ) and P‐STAT3 inhibitor C188‐9 (10 µ m ) for 48 h by WB and QPCR. ** p < 0.01, *** p < 0.001, **** p < 0.0001. (E,F) The protein and mRNA levels of NNMT and P‐STAT3 after treating detached cells with FAK inhibitor PF‐562271 (10 µ m for 48 h) and P‐STAT3 activator (1 µ m for 24 h) by WB and QPCR. ** p < 0.01, *** p < 0.001, **** p < 0.0001. (G,H) ChIP assay using P‐STAT3 antibodies in MDA‐MB‐231 cells cultured under suspension conditions was performed, followed by PCR and qPCR, which showed P‐STAT3 enrichment at the NNMT promoter vs. IgG control. ** p < 0.01. I: <t>Dual‐luciferase</t> reporter assays showed that co‐transfection with the full‐length NNMT promoter and a STAT3‐expressing plasmid significantly enhanced luciferase activity, whereas this enhancement was abolished when a truncated NNMT promoter was used. Data were presented as mean ± SEM. **** p < 0.0001.
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    Image Search Results


    Detachment‐induced NNMT is upregulated by the FAK‐STAT3 axis. (A) Representative image of the positive correlation between FAK and STAT3 in breast cancer and pan‐cancer according to the GEPIA 2 database analysis. (B) Representative results of STAT3, P‐STAT3, FAK, and P‐FAK proteins by WB in the detached BT‐549, MCF‐7, and MDA‐MB‐231 cells. (C,D) The protein and mRNA levels of NNMT and P‐STAT3 after treating detached cells with FAK inhibitor PF‐562271 (10 µ m ) and P‐STAT3 inhibitor C188‐9 (10 µ m ) for 48 h by WB and QPCR. ** p < 0.01, *** p < 0.001, **** p < 0.0001. (E,F) The protein and mRNA levels of NNMT and P‐STAT3 after treating detached cells with FAK inhibitor PF‐562271 (10 µ m for 48 h) and P‐STAT3 activator (1 µ m for 24 h) by WB and QPCR. ** p < 0.01, *** p < 0.001, **** p < 0.0001. (G,H) ChIP assay using P‐STAT3 antibodies in MDA‐MB‐231 cells cultured under suspension conditions was performed, followed by PCR and qPCR, which showed P‐STAT3 enrichment at the NNMT promoter vs. IgG control. ** p < 0.01. I: Dual‐luciferase reporter assays showed that co‐transfection with the full‐length NNMT promoter and a STAT3‐expressing plasmid significantly enhanced luciferase activity, whereas this enhancement was abolished when a truncated NNMT promoter was used. Data were presented as mean ± SEM. **** p < 0.0001.

    Journal: Advanced Science

    Article Title: Detachment‐Induced FAK‐STAT3‐NNMT Inhibits CTCs Anoikis to Promote Breast Cancer Metastasis by Enhancing Fatty Acid Oxidation

    doi: 10.1002/advs.202522837

    Figure Lengend Snippet: Detachment‐induced NNMT is upregulated by the FAK‐STAT3 axis. (A) Representative image of the positive correlation between FAK and STAT3 in breast cancer and pan‐cancer according to the GEPIA 2 database analysis. (B) Representative results of STAT3, P‐STAT3, FAK, and P‐FAK proteins by WB in the detached BT‐549, MCF‐7, and MDA‐MB‐231 cells. (C,D) The protein and mRNA levels of NNMT and P‐STAT3 after treating detached cells with FAK inhibitor PF‐562271 (10 µ m ) and P‐STAT3 inhibitor C188‐9 (10 µ m ) for 48 h by WB and QPCR. ** p < 0.01, *** p < 0.001, **** p < 0.0001. (E,F) The protein and mRNA levels of NNMT and P‐STAT3 after treating detached cells with FAK inhibitor PF‐562271 (10 µ m for 48 h) and P‐STAT3 activator (1 µ m for 24 h) by WB and QPCR. ** p < 0.01, *** p < 0.001, **** p < 0.0001. (G,H) ChIP assay using P‐STAT3 antibodies in MDA‐MB‐231 cells cultured under suspension conditions was performed, followed by PCR and qPCR, which showed P‐STAT3 enrichment at the NNMT promoter vs. IgG control. ** p < 0.01. I: Dual‐luciferase reporter assays showed that co‐transfection with the full‐length NNMT promoter and a STAT3‐expressing plasmid significantly enhanced luciferase activity, whereas this enhancement was abolished when a truncated NNMT promoter was used. Data were presented as mean ± SEM. **** p < 0.0001.

    Article Snippet: All plasmids used in this assay, including the firefly luciferase reporter vectors (e.g., pGL4‐basic containing the wild‐type or truncated NNMT promoter), the STAT3 expression plasmid, and the Renilla luciferase internal control plasmid (pRL‐TK), were custom‐synthesized by GeneChem Co., Ltd. (Shanghai, China).

    Techniques: Cell Culture, Suspension, Control, Luciferase, Cotransfection, Expressing, Plasmid Preparation, Activity Assay