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pgl-3 basic luciferase reporter  (Promega)

 
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    Structured Review

    Promega pgl-3 basic luciferase reporter
    Pgl 3 Basic Luciferase Reporter, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pgl-3+basic/pgl3+basic/pm39395526-115-28-30
    Average 90 stars, based on 1 article reviews
    pgl-3 basic luciferase reporter - by Bioz Stars, 2026-09
    90/100 stars

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    Related Articles

    Sequencing:

    Article Title: GATA2 Promotes Hematopoietic Development and Represses Cardiac Differentiation of Human Mesoderm
    Article Snippet: PITX2 and ISL1 promoter regions were amplified by PCR from human genomic DNA and inserted into the KpnI and NheI sites of pGL-3 Basic (Promega, Madison, WI) to generate reporter plasmids.

    Article Title: A circuit of COCH neurons encodes social-stress-induced anxiety via MTF1 activation of Cacna1h.
    Article Snippet: The sequence ofCacna1h 1020 promoter region was amplified by polymerase chain reaction(PCR) using mouse genomic DNA as a template with the primer set forward (MluI) (50-AT AGGTTGTCCAAGCAGTAAAGGG-30) and reverse (XhoI) (50-GGCTCACGTGTGCTCAGCC-30), and subcloned into the luciferase reporter vector pGL-3 basic (Promega).

    Article Title: Overexpression of homeodomain-interacting protein kinase 2 (HIPK2) attenuates sepsis-mediated liver injury by restoring autophagy
    Article Snippet: For further studies, primary hepatocytes were co-transfected with the internal control vector pmir-GLO (Promega) and pmir-GLO-HIPK2 3′-UTR using the Lipofectamine 2000 reagent (Thermo Fisher Scientific) for 20 h, followed by an incubation with the indicated chemicals for 10 h. Similarly, hepatocytes were co-transfected with the empty vector pGL-3 basic (Promega) and the pGL-3 vector containing the HIPK2 promoter using the Lipofectamine 2000 reagent for 20 h; the pRL vector was used as an internal control.

    Article Title: GATA2 Promotes Hematopoietic Development and Represses Cardiac Differentiation of Human Mesoderm
    Article Snippet: PITX2 and ISL1 promoter regions were amplified by PCR from human genomic DNA and inserted into the KpnI and NheI sites of pGL-3 Basic (Promega, Madison, WI) to generate reporter plasmids.

    Article Title: Identification of Synaptotagmin 10 as Effector of NPAS4-Mediated Protection from Excitotoxic Neurodegeneration
    Article Snippet: The Syt10 promoter fragments were amplified by PCR from rat genomic DNA as template using primers with the corresponding restriction sites for subcloning into the luciferase reporter vector pGL-3 basic (Promega; pGL3–4713 FW: 5′-GCGACGCGTAGTGATTTGTAGAAAAGACCACATGA-3′, MluI; pGL3–1036 FW: 5′-GCGACGCGTGGGGTAACTTTACCATAACCTGG-3′, MluI; pGL3–306 FW: 5′-GCGACGCGTTCAGGAAGGTGTGTGGGAA-3′, MluI; pGL3–306 RV: 5′-GCGGTCGACCTTGGCTTCTGCTCGCAG-3′, SalI; RR-I FW: 5′-GCGGGATCCTAAAGGGACAAGCTTGGGCT-3′, XhoI; RR-I RV: 5′-GCGGGTCGACAAAATTCTTGATAAATACTTGCTGAA 3′, BglII; RR-5′ FW: 5′-GCGGGTACCTCTTCAGGCTCAAAATTAGTGG-3′, KpnI; RR-5′ RV: 5′-GCGGGTACCCAAACTTCAAAAACATCATGCTA-3′, KpnI; RR-3′ FW: 5′-GTCGACGTTCTTCTCGCAGAGCCTCA-3′, SalI; RR-3′ RV: 5′-GTCGACAAGTCGGGGAAAGGGATTATT-3′, SalI; RR-1/-3 FW: 5′-GCGGGTACCAGAAGTTTACCCAGCTCACCA-3′, KpnI; RR-1/-2 RV: 5′-GCGGGTACCACTCACTGTGGTAAGCATTCATT-3′, KpnI; RR-2 FW: 5′-GCGGGTACCCTGCCCAGACTGTAGATAAACG-3′, KpnI; RR-3 RV: 5′-GCGGGTACCTTATCTACAGTCTGGGCAGAACA-3′, KpnI; exon 1 FW: 5′-GCGCTCGAGTACCTGTGCCACCTCCG-3′, XhoI; exon 1 RV: 5′-GCGAAGCTTAGTTTCCGCAAGGAGGAC-3′, HindIII).

    Article Title: Constitutive Activation of Nrf2 in Mice Expands Enterogenesis in Small Intestine Through Negative Regulation of Math1
    Article Snippet: The target PCR products directly cloned between the KpnI and NcoI sites of pGL-3 basic (Promega).

    Article Title: PPARγ attenuates hypoxia-induced hypertrophic transcriptional pathways in the heart
    Article Snippet: This NFAT-Luc construct was designed using nine copies of an NFAT binding site from the IL-4 promoter (5’-TGGAAAATT-3’) positioned 5’ to a minimal promoter from the alpha-myosin heavy chain gene (–164 to +16) and inserted upstream of the luciferase reporter in pGL-3 Basic (Promega Corporation, Madison, WI, USA) to create NFAT-luc.

    Article Title: Identification of Synaptotagmin 10 as Effector of NPAS4-Mediated Protection from Excitotoxic Neurodegeneration
    Article Snippet: The Syt10 promoter fragments were amplified by PCR from rat genomic DNA as template using primers with the corresponding restriction sites for subcloning into the luciferase reporter vector pGL-3 basic (Promega; pGL3–4713 FW: 5 -GCGACGCGTAGT GATTTGTAGAAAAGACCACATGA-3 , MluI; pGL3–1036 FW: 5 -GCGACGCGTGGGGTAACTTTACCATAACCTGG-3 , MluI; pGL3–306 FW: 5 -GCGACGCGTTCAGGAAGGTGTGTGGGAA-3 , MluI; pGL3– 306 RV: 5 -GCGGTCGACCTTGGCTTCTGCTCGCAG-3 , SalI; RR-I FW: 5 -GCGGGATCCTAAAGGGACAAGCTTGGGCT-3 , XhoI; RR-I RV: 5 - GCGGGTCGACAAAATTCTTGATAAATACTTGCTGAA 3 , BglII; RR-5 FW: 5 -GCGGGTACCTCTTCAGGCTCAAAATTAGTGG-3 , KpnI; RR-5 RV: 5 -GCGGGTACCCAAACTTCAAAAACATCATGCTA-3 , KpnI; RR-3 FW: 5 -GTCGACGTTCTTCTCGCAGAGCCTCA-3 , SalI; RR-3 RV: 5 -GTCGACAAGTCGGGGAAAGGGATTATT-3 , SalI; RR1/-3 FW: 5 -GCGGGTACCAGAAGTTTACCCAGCTCACCA-3 , KpnI; RR-1/-2 RV: 5 -GCGGGTACCACTCACTGTGGTAAGCATTCATT-3 , KpnI; RR-2 FW: 5 -GCGGGTACCCTGCCCAGACTGTAGATAAACG-3 , KpnI; RR-3 RV: 5 -GCGGGTACCTTATCTACAGTCTGGGCAGAACA3 , KpnI; exon 1 FW: 5 -GCGCTCGAGTACCTGTGCCACCTCCG-3 , XhoI; exon 1 RV: 5 -GCGAAGCTTAGTTTCCGCAAGGAGGAC-3 , HindIII).

    Amplification:

    Article Title: GATA2 Promotes Hematopoietic Development and Represses Cardiac Differentiation of Human Mesoderm
    Article Snippet: PITX2 and ISL1 promoter regions were amplified by PCR from human genomic DNA and inserted into the KpnI and NheI sites of pGL-3 Basic (Promega, Madison, WI) to generate reporter plasmids.

    Article Title: A circuit of COCH neurons encodes social-stress-induced anxiety via MTF1 activation of Cacna1h.
    Article Snippet: The sequence ofCacna1h 1020 promoter region was amplified by polymerase chain reaction(PCR) using mouse genomic DNA as a template with the primer set forward (MluI) (50-AT AGGTTGTCCAAGCAGTAAAGGG-30) and reverse (XhoI) (50-GGCTCACGTGTGCTCAGCC-30), and subcloned into the luciferase reporter vector pGL-3 basic (Promega).

    Article Title: Overexpression of homeodomain-interacting protein kinase 2 (HIPK2) attenuates sepsis-mediated liver injury by restoring autophagy
    Article Snippet: For further studies, primary hepatocytes were co-transfected with the internal control vector pmir-GLO (Promega) and pmir-GLO-HIPK2 3′-UTR using the Lipofectamine 2000 reagent (Thermo Fisher Scientific) for 20 h, followed by an incubation with the indicated chemicals for 10 h. Similarly, hepatocytes were co-transfected with the empty vector pGL-3 basic (Promega) and the pGL-3 vector containing the HIPK2 promoter using the Lipofectamine 2000 reagent for 20 h; the pRL vector was used as an internal control.

    Article Title: GATA2 Promotes Hematopoietic Development and Represses Cardiac Differentiation of Human Mesoderm
    Article Snippet: PITX2 and ISL1 promoter regions were amplified by PCR from human genomic DNA and inserted into the KpnI and NheI sites of pGL-3 Basic (Promega, Madison, WI) to generate reporter plasmids.

    Article Title: Identification of Synaptotagmin 10 as Effector of NPAS4-Mediated Protection from Excitotoxic Neurodegeneration
    Article Snippet: The Syt10 promoter fragments were amplified by PCR from rat genomic DNA as template using primers with the corresponding restriction sites for subcloning into the luciferase reporter vector pGL-3 basic (Promega; pGL3–4713 FW: 5′-GCGACGCGTAGTGATTTGTAGAAAAGACCACATGA-3′, MluI; pGL3–1036 FW: 5′-GCGACGCGTGGGGTAACTTTACCATAACCTGG-3′, MluI; pGL3–306 FW: 5′-GCGACGCGTTCAGGAAGGTGTGTGGGAA-3′, MluI; pGL3–306 RV: 5′-GCGGTCGACCTTGGCTTCTGCTCGCAG-3′, SalI; RR-I FW: 5′-GCGGGATCCTAAAGGGACAAGCTTGGGCT-3′, XhoI; RR-I RV: 5′-GCGGGTCGACAAAATTCTTGATAAATACTTGCTGAA 3′, BglII; RR-5′ FW: 5′-GCGGGTACCTCTTCAGGCTCAAAATTAGTGG-3′, KpnI; RR-5′ RV: 5′-GCGGGTACCCAAACTTCAAAAACATCATGCTA-3′, KpnI; RR-3′ FW: 5′-GTCGACGTTCTTCTCGCAGAGCCTCA-3′, SalI; RR-3′ RV: 5′-GTCGACAAGTCGGGGAAAGGGATTATT-3′, SalI; RR-1/-3 FW: 5′-GCGGGTACCAGAAGTTTACCCAGCTCACCA-3′, KpnI; RR-1/-2 RV: 5′-GCGGGTACCACTCACTGTGGTAAGCATTCATT-3′, KpnI; RR-2 FW: 5′-GCGGGTACCCTGCCCAGACTGTAGATAAACG-3′, KpnI; RR-3 RV: 5′-GCGGGTACCTTATCTACAGTCTGGGCAGAACA-3′, KpnI; exon 1 FW: 5′-GCGCTCGAGTACCTGTGCCACCTCCG-3′, XhoI; exon 1 RV: 5′-GCGAAGCTTAGTTTCCGCAAGGAGGAC-3′, HindIII).

    Article Title: Constitutive Activation of Nrf2 in Mice Expands Enterogenesis in Small Intestine Through Negative Regulation of Math1
    Article Snippet: The target PCR products directly cloned between the KpnI and NcoI sites of pGL-3 basic (Promega).

    Article Title: PPARγ attenuates hypoxia-induced hypertrophic transcriptional pathways in the heart
    Article Snippet: This NFAT-Luc construct was designed using nine copies of an NFAT binding site from the IL-4 promoter (5’-TGGAAAATT-3’) positioned 5’ to a minimal promoter from the alpha-myosin heavy chain gene (–164 to +16) and inserted upstream of the luciferase reporter in pGL-3 Basic (Promega Corporation, Madison, WI, USA) to create NFAT-luc.

    Article Title: Identification of Synaptotagmin 10 as Effector of NPAS4-Mediated Protection from Excitotoxic Neurodegeneration
    Article Snippet: The Syt10 promoter fragments were amplified by PCR from rat genomic DNA as template using primers with the corresponding restriction sites for subcloning into the luciferase reporter vector pGL-3 basic (Promega; pGL3–4713 FW: 5 -GCGACGCGTAGT GATTTGTAGAAAAGACCACATGA-3 , MluI; pGL3–1036 FW: 5 -GCGACGCGTGGGGTAACTTTACCATAACCTGG-3 , MluI; pGL3–306 FW: 5 -GCGACGCGTTCAGGAAGGTGTGTGGGAA-3 , MluI; pGL3– 306 RV: 5 -GCGGTCGACCTTGGCTTCTGCTCGCAG-3 , SalI; RR-I FW: 5 -GCGGGATCCTAAAGGGACAAGCTTGGGCT-3 , XhoI; RR-I RV: 5 - GCGGGTCGACAAAATTCTTGATAAATACTTGCTGAA 3 , BglII; RR-5 FW: 5 -GCGGGTACCTCTTCAGGCTCAAAATTAGTGG-3 , KpnI; RR-5 RV: 5 -GCGGGTACCCAAACTTCAAAAACATCATGCTA-3 , KpnI; RR-3 FW: 5 -GTCGACGTTCTTCTCGCAGAGCCTCA-3 , SalI; RR-3 RV: 5 -GTCGACAAGTCGGGGAAAGGGATTATT-3 , SalI; RR1/-3 FW: 5 -GCGGGTACCAGAAGTTTACCCAGCTCACCA-3 , KpnI; RR-1/-2 RV: 5 -GCGGGTACCACTCACTGTGGTAAGCATTCATT-3 , KpnI; RR-2 FW: 5 -GCGGGTACCCTGCCCAGACTGTAGATAAACG-3 , KpnI; RR-3 RV: 5 -GCGGGTACCTTATCTACAGTCTGGGCAGAACA3 , KpnI; exon 1 FW: 5 -GCGCTCGAGTACCTGTGCCACCTCCG-3 , XhoI; exon 1 RV: 5 -GCGAAGCTTAGTTTCCGCAAGGAGGAC-3 , HindIII).

    Polymerase Chain Reaction:

    Article Title: GATA2 Promotes Hematopoietic Development and Represses Cardiac Differentiation of Human Mesoderm
    Article Snippet: PITX2 and ISL1 promoter regions were amplified by PCR from human genomic DNA and inserted into the KpnI and NheI sites of pGL-3 Basic (Promega, Madison, WI) to generate reporter plasmids.

    Article Title: A circuit of COCH neurons encodes social-stress-induced anxiety via MTF1 activation of Cacna1h.
    Article Snippet: The sequence ofCacna1h 1020 promoter region was amplified by polymerase chain reaction(PCR) using mouse genomic DNA as a template with the primer set forward (MluI) (50-AT AGGTTGTCCAAGCAGTAAAGGG-30) and reverse (XhoI) (50-GGCTCACGTGTGCTCAGCC-30), and subcloned into the luciferase reporter vector pGL-3 basic (Promega).

    Article Title: Overexpression of homeodomain-interacting protein kinase 2 (HIPK2) attenuates sepsis-mediated liver injury by restoring autophagy
    Article Snippet: For further studies, primary hepatocytes were co-transfected with the internal control vector pmir-GLO (Promega) and pmir-GLO-HIPK2 3′-UTR using the Lipofectamine 2000 reagent (Thermo Fisher Scientific) for 20 h, followed by an incubation with the indicated chemicals for 10 h. Similarly, hepatocytes were co-transfected with the empty vector pGL-3 basic (Promega) and the pGL-3 vector containing the HIPK2 promoter using the Lipofectamine 2000 reagent for 20 h; the pRL vector was used as an internal control.

    Article Title: GATA2 Promotes Hematopoietic Development and Represses Cardiac Differentiation of Human Mesoderm
    Article Snippet: PITX2 and ISL1 promoter regions were amplified by PCR from human genomic DNA and inserted into the KpnI and NheI sites of pGL-3 Basic (Promega, Madison, WI) to generate reporter plasmids.

    Article Title: Identification of Synaptotagmin 10 as Effector of NPAS4-Mediated Protection from Excitotoxic Neurodegeneration
    Article Snippet: The Syt10 promoter fragments were amplified by PCR from rat genomic DNA as template using primers with the corresponding restriction sites for subcloning into the luciferase reporter vector pGL-3 basic (Promega; pGL3–4713 FW: 5′-GCGACGCGTAGTGATTTGTAGAAAAGACCACATGA-3′, MluI; pGL3–1036 FW: 5′-GCGACGCGTGGGGTAACTTTACCATAACCTGG-3′, MluI; pGL3–306 FW: 5′-GCGACGCGTTCAGGAAGGTGTGTGGGAA-3′, MluI; pGL3–306 RV: 5′-GCGGTCGACCTTGGCTTCTGCTCGCAG-3′, SalI; RR-I FW: 5′-GCGGGATCCTAAAGGGACAAGCTTGGGCT-3′, XhoI; RR-I RV: 5′-GCGGGTCGACAAAATTCTTGATAAATACTTGCTGAA 3′, BglII; RR-5′ FW: 5′-GCGGGTACCTCTTCAGGCTCAAAATTAGTGG-3′, KpnI; RR-5′ RV: 5′-GCGGGTACCCAAACTTCAAAAACATCATGCTA-3′, KpnI; RR-3′ FW: 5′-GTCGACGTTCTTCTCGCAGAGCCTCA-3′, SalI; RR-3′ RV: 5′-GTCGACAAGTCGGGGAAAGGGATTATT-3′, SalI; RR-1/-3 FW: 5′-GCGGGTACCAGAAGTTTACCCAGCTCACCA-3′, KpnI; RR-1/-2 RV: 5′-GCGGGTACCACTCACTGTGGTAAGCATTCATT-3′, KpnI; RR-2 FW: 5′-GCGGGTACCCTGCCCAGACTGTAGATAAACG-3′, KpnI; RR-3 RV: 5′-GCGGGTACCTTATCTACAGTCTGGGCAGAACA-3′, KpnI; exon 1 FW: 5′-GCGCTCGAGTACCTGTGCCACCTCCG-3′, XhoI; exon 1 RV: 5′-GCGAAGCTTAGTTTCCGCAAGGAGGAC-3′, HindIII).

    Article Title: Constitutive Activation of Nrf2 in Mice Expands Enterogenesis in Small Intestine Through Negative Regulation of Math1
    Article Snippet: The target PCR products directly cloned between the KpnI and NcoI sites of pGL-3 basic (Promega).

    Article Title: PPARγ attenuates hypoxia-induced hypertrophic transcriptional pathways in the heart
    Article Snippet: This NFAT-Luc construct was designed using nine copies of an NFAT binding site from the IL-4 promoter (5’-TGGAAAATT-3’) positioned 5’ to a minimal promoter from the alpha-myosin heavy chain gene (–164 to +16) and inserted upstream of the luciferase reporter in pGL-3 Basic (Promega Corporation, Madison, WI, USA) to create NFAT-luc.

    Article Title: Identification of Synaptotagmin 10 as Effector of NPAS4-Mediated Protection from Excitotoxic Neurodegeneration
    Article Snippet: The Syt10 promoter fragments were amplified by PCR from rat genomic DNA as template using primers with the corresponding restriction sites for subcloning into the luciferase reporter vector pGL-3 basic (Promega; pGL3–4713 FW: 5 -GCGACGCGTAGT GATTTGTAGAAAAGACCACATGA-3 , MluI; pGL3–1036 FW: 5 -GCGACGCGTGGGGTAACTTTACCATAACCTGG-3 , MluI; pGL3–306 FW: 5 -GCGACGCGTTCAGGAAGGTGTGTGGGAA-3 , MluI; pGL3– 306 RV: 5 -GCGGTCGACCTTGGCTTCTGCTCGCAG-3 , SalI; RR-I FW: 5 -GCGGGATCCTAAAGGGACAAGCTTGGGCT-3 , XhoI; RR-I RV: 5 - GCGGGTCGACAAAATTCTTGATAAATACTTGCTGAA 3 , BglII; RR-5 FW: 5 -GCGGGTACCTCTTCAGGCTCAAAATTAGTGG-3 , KpnI; RR-5 RV: 5 -GCGGGTACCCAAACTTCAAAAACATCATGCTA-3 , KpnI; RR-3 FW: 5 -GTCGACGTTCTTCTCGCAGAGCCTCA-3 , SalI; RR-3 RV: 5 -GTCGACAAGTCGGGGAAAGGGATTATT-3 , SalI; RR1/-3 FW: 5 -GCGGGTACCAGAAGTTTACCCAGCTCACCA-3 , KpnI; RR-1/-2 RV: 5 -GCGGGTACCACTCACTGTGGTAAGCATTCATT-3 , KpnI; RR-2 FW: 5 -GCGGGTACCCTGCCCAGACTGTAGATAAACG-3 , KpnI; RR-3 RV: 5 -GCGGGTACCTTATCTACAGTCTGGGCAGAACA3 , KpnI; exon 1 FW: 5 -GCGCTCGAGTACCTGTGCCACCTCCG-3 , XhoI; exon 1 RV: 5 -GCGAAGCTTAGTTTCCGCAAGGAGGAC-3 , HindIII).

    Luciferase:

    Article Title: GATA2 Promotes Hematopoietic Development and Represses Cardiac Differentiation of Human Mesoderm
    Article Snippet: PITX2 and ISL1 promoter regions were amplified by PCR from human genomic DNA and inserted into the KpnI and NheI sites of pGL-3 Basic (Promega, Madison, WI) to generate reporter plasmids.

    Article Title: A circuit of COCH neurons encodes social-stress-induced anxiety via MTF1 activation of Cacna1h.
    Article Snippet: The sequence ofCacna1h 1020 promoter region was amplified by polymerase chain reaction(PCR) using mouse genomic DNA as a template with the primer set forward (MluI) (50-AT AGGTTGTCCAAGCAGTAAAGGG-30) and reverse (XhoI) (50-GGCTCACGTGTGCTCAGCC-30), and subcloned into the luciferase reporter vector pGL-3 basic (Promega).

    Article Title: Overexpression of homeodomain-interacting protein kinase 2 (HIPK2) attenuates sepsis-mediated liver injury by restoring autophagy
    Article Snippet: For further studies, primary hepatocytes were co-transfected with the internal control vector pmir-GLO (Promega) and pmir-GLO-HIPK2 3′-UTR using the Lipofectamine 2000 reagent (Thermo Fisher Scientific) for 20 h, followed by an incubation with the indicated chemicals for 10 h. Similarly, hepatocytes were co-transfected with the empty vector pGL-3 basic (Promega) and the pGL-3 vector containing the HIPK2 promoter using the Lipofectamine 2000 reagent for 20 h; the pRL vector was used as an internal control.

    Article Title: GATA2 Promotes Hematopoietic Development and Represses Cardiac Differentiation of Human Mesoderm
    Article Snippet: PITX2 and ISL1 promoter regions were amplified by PCR from human genomic DNA and inserted into the KpnI and NheI sites of pGL-3 Basic (Promega, Madison, WI) to generate reporter plasmids.

    Article Title: Identification of Synaptotagmin 10 as Effector of NPAS4-Mediated Protection from Excitotoxic Neurodegeneration
    Article Snippet: The Syt10 promoter fragments were amplified by PCR from rat genomic DNA as template using primers with the corresponding restriction sites for subcloning into the luciferase reporter vector pGL-3 basic (Promega; pGL3–4713 FW: 5′-GCGACGCGTAGTGATTTGTAGAAAAGACCACATGA-3′, MluI; pGL3–1036 FW: 5′-GCGACGCGTGGGGTAACTTTACCATAACCTGG-3′, MluI; pGL3–306 FW: 5′-GCGACGCGTTCAGGAAGGTGTGTGGGAA-3′, MluI; pGL3–306 RV: 5′-GCGGTCGACCTTGGCTTCTGCTCGCAG-3′, SalI; RR-I FW: 5′-GCGGGATCCTAAAGGGACAAGCTTGGGCT-3′, XhoI; RR-I RV: 5′-GCGGGTCGACAAAATTCTTGATAAATACTTGCTGAA 3′, BglII; RR-5′ FW: 5′-GCGGGTACCTCTTCAGGCTCAAAATTAGTGG-3′, KpnI; RR-5′ RV: 5′-GCGGGTACCCAAACTTCAAAAACATCATGCTA-3′, KpnI; RR-3′ FW: 5′-GTCGACGTTCTTCTCGCAGAGCCTCA-3′, SalI; RR-3′ RV: 5′-GTCGACAAGTCGGGGAAAGGGATTATT-3′, SalI; RR-1/-3 FW: 5′-GCGGGTACCAGAAGTTTACCCAGCTCACCA-3′, KpnI; RR-1/-2 RV: 5′-GCGGGTACCACTCACTGTGGTAAGCATTCATT-3′, KpnI; RR-2 FW: 5′-GCGGGTACCCTGCCCAGACTGTAGATAAACG-3′, KpnI; RR-3 RV: 5′-GCGGGTACCTTATCTACAGTCTGGGCAGAACA-3′, KpnI; exon 1 FW: 5′-GCGCTCGAGTACCTGTGCCACCTCCG-3′, XhoI; exon 1 RV: 5′-GCGAAGCTTAGTTTCCGCAAGGAGGAC-3′, HindIII).

    Article Title: Constitutive Activation of Nrf2 in Mice Expands Enterogenesis in Small Intestine Through Negative Regulation of Math1
    Article Snippet: The target PCR products directly cloned between the KpnI and NcoI sites of pGL-3 basic (Promega).

    Article Title: PPARγ attenuates hypoxia-induced hypertrophic transcriptional pathways in the heart
    Article Snippet: This NFAT-Luc construct was designed using nine copies of an NFAT binding site from the IL-4 promoter (5’-TGGAAAATT-3’) positioned 5’ to a minimal promoter from the alpha-myosin heavy chain gene (–164 to +16) and inserted upstream of the luciferase reporter in pGL-3 Basic (Promega Corporation, Madison, WI, USA) to create NFAT-luc.

    Article Title: Identification of Synaptotagmin 10 as Effector of NPAS4-Mediated Protection from Excitotoxic Neurodegeneration
    Article Snippet: The Syt10 promoter fragments were amplified by PCR from rat genomic DNA as template using primers with the corresponding restriction sites for subcloning into the luciferase reporter vector pGL-3 basic (Promega; pGL3–4713 FW: 5 -GCGACGCGTAGT GATTTGTAGAAAAGACCACATGA-3 , MluI; pGL3–1036 FW: 5 -GCGACGCGTGGGGTAACTTTACCATAACCTGG-3 , MluI; pGL3–306 FW: 5 -GCGACGCGTTCAGGAAGGTGTGTGGGAA-3 , MluI; pGL3– 306 RV: 5 -GCGGTCGACCTTGGCTTCTGCTCGCAG-3 , SalI; RR-I FW: 5 -GCGGGATCCTAAAGGGACAAGCTTGGGCT-3 , XhoI; RR-I RV: 5 - GCGGGTCGACAAAATTCTTGATAAATACTTGCTGAA 3 , BglII; RR-5 FW: 5 -GCGGGTACCTCTTCAGGCTCAAAATTAGTGG-3 , KpnI; RR-5 RV: 5 -GCGGGTACCCAAACTTCAAAAACATCATGCTA-3 , KpnI; RR-3 FW: 5 -GTCGACGTTCTTCTCGCAGAGCCTCA-3 , SalI; RR-3 RV: 5 -GTCGACAAGTCGGGGAAAGGGATTATT-3 , SalI; RR1/-3 FW: 5 -GCGGGTACCAGAAGTTTACCCAGCTCACCA-3 , KpnI; RR-1/-2 RV: 5 -GCGGGTACCACTCACTGTGGTAAGCATTCATT-3 , KpnI; RR-2 FW: 5 -GCGGGTACCCTGCCCAGACTGTAGATAAACG-3 , KpnI; RR-3 RV: 5 -GCGGGTACCTTATCTACAGTCTGGGCAGAACA3 , KpnI; exon 1 FW: 5 -GCGCTCGAGTACCTGTGCCACCTCCG-3 , XhoI; exon 1 RV: 5 -GCGAAGCTTAGTTTCCGCAAGGAGGAC-3 , HindIII).

    Plasmid Preparation:

    Article Title: GATA2 Promotes Hematopoietic Development and Represses Cardiac Differentiation of Human Mesoderm
    Article Snippet: PITX2 and ISL1 promoter regions were amplified by PCR from human genomic DNA and inserted into the KpnI and NheI sites of pGL-3 Basic (Promega, Madison, WI) to generate reporter plasmids.

    Article Title: A circuit of COCH neurons encodes social-stress-induced anxiety via MTF1 activation of Cacna1h.
    Article Snippet: The sequence ofCacna1h 1020 promoter region was amplified by polymerase chain reaction(PCR) using mouse genomic DNA as a template with the primer set forward (MluI) (50-AT AGGTTGTCCAAGCAGTAAAGGG-30) and reverse (XhoI) (50-GGCTCACGTGTGCTCAGCC-30), and subcloned into the luciferase reporter vector pGL-3 basic (Promega).

    Article Title: Overexpression of homeodomain-interacting protein kinase 2 (HIPK2) attenuates sepsis-mediated liver injury by restoring autophagy
    Article Snippet: For further studies, primary hepatocytes were co-transfected with the internal control vector pmir-GLO (Promega) and pmir-GLO-HIPK2 3′-UTR using the Lipofectamine 2000 reagent (Thermo Fisher Scientific) for 20 h, followed by an incubation with the indicated chemicals for 10 h. Similarly, hepatocytes were co-transfected with the empty vector pGL-3 basic (Promega) and the pGL-3 vector containing the HIPK2 promoter using the Lipofectamine 2000 reagent for 20 h; the pRL vector was used as an internal control.

    Article Title: GATA2 Promotes Hematopoietic Development and Represses Cardiac Differentiation of Human Mesoderm
    Article Snippet: PITX2 and ISL1 promoter regions were amplified by PCR from human genomic DNA and inserted into the KpnI and NheI sites of pGL-3 Basic (Promega, Madison, WI) to generate reporter plasmids.

    Article Title: Identification of Synaptotagmin 10 as Effector of NPAS4-Mediated Protection from Excitotoxic Neurodegeneration
    Article Snippet: The Syt10 promoter fragments were amplified by PCR from rat genomic DNA as template using primers with the corresponding restriction sites for subcloning into the luciferase reporter vector pGL-3 basic (Promega; pGL3–4713 FW: 5′-GCGACGCGTAGTGATTTGTAGAAAAGACCACATGA-3′, MluI; pGL3–1036 FW: 5′-GCGACGCGTGGGGTAACTTTACCATAACCTGG-3′, MluI; pGL3–306 FW: 5′-GCGACGCGTTCAGGAAGGTGTGTGGGAA-3′, MluI; pGL3–306 RV: 5′-GCGGTCGACCTTGGCTTCTGCTCGCAG-3′, SalI; RR-I FW: 5′-GCGGGATCCTAAAGGGACAAGCTTGGGCT-3′, XhoI; RR-I RV: 5′-GCGGGTCGACAAAATTCTTGATAAATACTTGCTGAA 3′, BglII; RR-5′ FW: 5′-GCGGGTACCTCTTCAGGCTCAAAATTAGTGG-3′, KpnI; RR-5′ RV: 5′-GCGGGTACCCAAACTTCAAAAACATCATGCTA-3′, KpnI; RR-3′ FW: 5′-GTCGACGTTCTTCTCGCAGAGCCTCA-3′, SalI; RR-3′ RV: 5′-GTCGACAAGTCGGGGAAAGGGATTATT-3′, SalI; RR-1/-3 FW: 5′-GCGGGTACCAGAAGTTTACCCAGCTCACCA-3′, KpnI; RR-1/-2 RV: 5′-GCGGGTACCACTCACTGTGGTAAGCATTCATT-3′, KpnI; RR-2 FW: 5′-GCGGGTACCCTGCCCAGACTGTAGATAAACG-3′, KpnI; RR-3 RV: 5′-GCGGGTACCTTATCTACAGTCTGGGCAGAACA-3′, KpnI; exon 1 FW: 5′-GCGCTCGAGTACCTGTGCCACCTCCG-3′, XhoI; exon 1 RV: 5′-GCGAAGCTTAGTTTCCGCAAGGAGGAC-3′, HindIII).

    Article Title: Constitutive Activation of Nrf2 in Mice Expands Enterogenesis in Small Intestine Through Negative Regulation of Math1
    Article Snippet: The target PCR products directly cloned between the KpnI and NcoI sites of pGL-3 basic (Promega).

    Article Title: PPARγ attenuates hypoxia-induced hypertrophic transcriptional pathways in the heart
    Article Snippet: This NFAT-Luc construct was designed using nine copies of an NFAT binding site from the IL-4 promoter (5’-TGGAAAATT-3’) positioned 5’ to a minimal promoter from the alpha-myosin heavy chain gene (–164 to +16) and inserted upstream of the luciferase reporter in pGL-3 Basic (Promega Corporation, Madison, WI, USA) to create NFAT-luc.

    Article Title: Identification of Synaptotagmin 10 as Effector of NPAS4-Mediated Protection from Excitotoxic Neurodegeneration
    Article Snippet: The Syt10 promoter fragments were amplified by PCR from rat genomic DNA as template using primers with the corresponding restriction sites for subcloning into the luciferase reporter vector pGL-3 basic (Promega; pGL3–4713 FW: 5 -GCGACGCGTAGT GATTTGTAGAAAAGACCACATGA-3 , MluI; pGL3–1036 FW: 5 -GCGACGCGTGGGGTAACTTTACCATAACCTGG-3 , MluI; pGL3–306 FW: 5 -GCGACGCGTTCAGGAAGGTGTGTGGGAA-3 , MluI; pGL3– 306 RV: 5 -GCGGTCGACCTTGGCTTCTGCTCGCAG-3 , SalI; RR-I FW: 5 -GCGGGATCCTAAAGGGACAAGCTTGGGCT-3 , XhoI; RR-I RV: 5 - GCGGGTCGACAAAATTCTTGATAAATACTTGCTGAA 3 , BglII; RR-5 FW: 5 -GCGGGTACCTCTTCAGGCTCAAAATTAGTGG-3 , KpnI; RR-5 RV: 5 -GCGGGTACCCAAACTTCAAAAACATCATGCTA-3 , KpnI; RR-3 FW: 5 -GTCGACGTTCTTCTCGCAGAGCCTCA-3 , SalI; RR-3 RV: 5 -GTCGACAAGTCGGGGAAAGGGATTATT-3 , SalI; RR1/-3 FW: 5 -GCGGGTACCAGAAGTTTACCCAGCTCACCA-3 , KpnI; RR-1/-2 RV: 5 -GCGGGTACCACTCACTGTGGTAAGCATTCATT-3 , KpnI; RR-2 FW: 5 -GCGGGTACCCTGCCCAGACTGTAGATAAACG-3 , KpnI; RR-3 RV: 5 -GCGGGTACCTTATCTACAGTCTGGGCAGAACA3 , KpnI; exon 1 FW: 5 -GCGCTCGAGTACCTGTGCCACCTCCG-3 , XhoI; exon 1 RV: 5 -GCGAAGCTTAGTTTCCGCAAGGAGGAC-3 , HindIII).

    Clone Assay:

    Article Title: GATA2 Promotes Hematopoietic Development and Represses Cardiac Differentiation of Human Mesoderm
    Article Snippet: PITX2 and ISL1 promoter regions were amplified by PCR from human genomic DNA and inserted into the KpnI and NheI sites of pGL-3 Basic (Promega, Madison, WI) to generate reporter plasmids.

    Article Title: A circuit of COCH neurons encodes social-stress-induced anxiety via MTF1 activation of Cacna1h.
    Article Snippet: The sequence ofCacna1h 1020 promoter region was amplified by polymerase chain reaction(PCR) using mouse genomic DNA as a template with the primer set forward (MluI) (50-AT AGGTTGTCCAAGCAGTAAAGGG-30) and reverse (XhoI) (50-GGCTCACGTGTGCTCAGCC-30), and subcloned into the luciferase reporter vector pGL-3 basic (Promega).

    Article Title: Overexpression of homeodomain-interacting protein kinase 2 (HIPK2) attenuates sepsis-mediated liver injury by restoring autophagy
    Article Snippet: For further studies, primary hepatocytes were co-transfected with the internal control vector pmir-GLO (Promega) and pmir-GLO-HIPK2 3′-UTR using the Lipofectamine 2000 reagent (Thermo Fisher Scientific) for 20 h, followed by an incubation with the indicated chemicals for 10 h. Similarly, hepatocytes were co-transfected with the empty vector pGL-3 basic (Promega) and the pGL-3 vector containing the HIPK2 promoter using the Lipofectamine 2000 reagent for 20 h; the pRL vector was used as an internal control.

    Article Title: GATA2 Promotes Hematopoietic Development and Represses Cardiac Differentiation of Human Mesoderm
    Article Snippet: PITX2 and ISL1 promoter regions were amplified by PCR from human genomic DNA and inserted into the KpnI and NheI sites of pGL-3 Basic (Promega, Madison, WI) to generate reporter plasmids.

    Article Title: Identification of Synaptotagmin 10 as Effector of NPAS4-Mediated Protection from Excitotoxic Neurodegeneration
    Article Snippet: The Syt10 promoter fragments were amplified by PCR from rat genomic DNA as template using primers with the corresponding restriction sites for subcloning into the luciferase reporter vector pGL-3 basic (Promega; pGL3–4713 FW: 5′-GCGACGCGTAGTGATTTGTAGAAAAGACCACATGA-3′, MluI; pGL3–1036 FW: 5′-GCGACGCGTGGGGTAACTTTACCATAACCTGG-3′, MluI; pGL3–306 FW: 5′-GCGACGCGTTCAGGAAGGTGTGTGGGAA-3′, MluI; pGL3–306 RV: 5′-GCGGTCGACCTTGGCTTCTGCTCGCAG-3′, SalI; RR-I FW: 5′-GCGGGATCCTAAAGGGACAAGCTTGGGCT-3′, XhoI; RR-I RV: 5′-GCGGGTCGACAAAATTCTTGATAAATACTTGCTGAA 3′, BglII; RR-5′ FW: 5′-GCGGGTACCTCTTCAGGCTCAAAATTAGTGG-3′, KpnI; RR-5′ RV: 5′-GCGGGTACCCAAACTTCAAAAACATCATGCTA-3′, KpnI; RR-3′ FW: 5′-GTCGACGTTCTTCTCGCAGAGCCTCA-3′, SalI; RR-3′ RV: 5′-GTCGACAAGTCGGGGAAAGGGATTATT-3′, SalI; RR-1/-3 FW: 5′-GCGGGTACCAGAAGTTTACCCAGCTCACCA-3′, KpnI; RR-1/-2 RV: 5′-GCGGGTACCACTCACTGTGGTAAGCATTCATT-3′, KpnI; RR-2 FW: 5′-GCGGGTACCCTGCCCAGACTGTAGATAAACG-3′, KpnI; RR-3 RV: 5′-GCGGGTACCTTATCTACAGTCTGGGCAGAACA-3′, KpnI; exon 1 FW: 5′-GCGCTCGAGTACCTGTGCCACCTCCG-3′, XhoI; exon 1 RV: 5′-GCGAAGCTTAGTTTCCGCAAGGAGGAC-3′, HindIII).

    Article Title: Constitutive Activation of Nrf2 in Mice Expands Enterogenesis in Small Intestine Through Negative Regulation of Math1
    Article Snippet: The target PCR products directly cloned between the KpnI and NcoI sites of pGL-3 basic (Promega).

    Article Title: PPARγ attenuates hypoxia-induced hypertrophic transcriptional pathways in the heart
    Article Snippet: This NFAT-Luc construct was designed using nine copies of an NFAT binding site from the IL-4 promoter (5’-TGGAAAATT-3’) positioned 5’ to a minimal promoter from the alpha-myosin heavy chain gene (–164 to +16) and inserted upstream of the luciferase reporter in pGL-3 Basic (Promega Corporation, Madison, WI, USA) to create NFAT-luc.

    Article Title: Identification of Synaptotagmin 10 as Effector of NPAS4-Mediated Protection from Excitotoxic Neurodegeneration
    Article Snippet: The Syt10 promoter fragments were amplified by PCR from rat genomic DNA as template using primers with the corresponding restriction sites for subcloning into the luciferase reporter vector pGL-3 basic (Promega; pGL3–4713 FW: 5 -GCGACGCGTAGT GATTTGTAGAAAAGACCACATGA-3 , MluI; pGL3–1036 FW: 5 -GCGACGCGTGGGGTAACTTTACCATAACCTGG-3 , MluI; pGL3–306 FW: 5 -GCGACGCGTTCAGGAAGGTGTGTGGGAA-3 , MluI; pGL3– 306 RV: 5 -GCGGTCGACCTTGGCTTCTGCTCGCAG-3 , SalI; RR-I FW: 5 -GCGGGATCCTAAAGGGACAAGCTTGGGCT-3 , XhoI; RR-I RV: 5 - GCGGGTCGACAAAATTCTTGATAAATACTTGCTGAA 3 , BglII; RR-5 FW: 5 -GCGGGTACCTCTTCAGGCTCAAAATTAGTGG-3 , KpnI; RR-5 RV: 5 -GCGGGTACCCAAACTTCAAAAACATCATGCTA-3 , KpnI; RR-3 FW: 5 -GTCGACGTTCTTCTCGCAGAGCCTCA-3 , SalI; RR-3 RV: 5 -GTCGACAAGTCGGGGAAAGGGATTATT-3 , SalI; RR1/-3 FW: 5 -GCGGGTACCAGAAGTTTACCCAGCTCACCA-3 , KpnI; RR-1/-2 RV: 5 -GCGGGTACCACTCACTGTGGTAAGCATTCATT-3 , KpnI; RR-2 FW: 5 -GCGGGTACCCTGCCCAGACTGTAGATAAACG-3 , KpnI; RR-3 RV: 5 -GCGGGTACCTTATCTACAGTCTGGGCAGAACA3 , KpnI; exon 1 FW: 5 -GCGCTCGAGTACCTGTGCCACCTCCG-3 , XhoI; exon 1 RV: 5 -GCGAAGCTTAGTTTCCGCAAGGAGGAC-3 , HindIII).

    Incubation:

    Article Title: GATA2 Promotes Hematopoietic Development and Represses Cardiac Differentiation of Human Mesoderm
    Article Snippet: PITX2 and ISL1 promoter regions were amplified by PCR from human genomic DNA and inserted into the KpnI and NheI sites of pGL-3 Basic (Promega, Madison, WI) to generate reporter plasmids.

    Article Title: A circuit of COCH neurons encodes social-stress-induced anxiety via MTF1 activation of Cacna1h.
    Article Snippet: The sequence ofCacna1h 1020 promoter region was amplified by polymerase chain reaction(PCR) using mouse genomic DNA as a template with the primer set forward (MluI) (50-AT AGGTTGTCCAAGCAGTAAAGGG-30) and reverse (XhoI) (50-GGCTCACGTGTGCTCAGCC-30), and subcloned into the luciferase reporter vector pGL-3 basic (Promega).

    Article Title: Overexpression of homeodomain-interacting protein kinase 2 (HIPK2) attenuates sepsis-mediated liver injury by restoring autophagy
    Article Snippet: For further studies, primary hepatocytes were co-transfected with the internal control vector pmir-GLO (Promega) and pmir-GLO-HIPK2 3′-UTR using the Lipofectamine 2000 reagent (Thermo Fisher Scientific) for 20 h, followed by an incubation with the indicated chemicals for 10 h. Similarly, hepatocytes were co-transfected with the empty vector pGL-3 basic (Promega) and the pGL-3 vector containing the HIPK2 promoter using the Lipofectamine 2000 reagent for 20 h; the pRL vector was used as an internal control.

    Article Title: GATA2 Promotes Hematopoietic Development and Represses Cardiac Differentiation of Human Mesoderm
    Article Snippet: PITX2 and ISL1 promoter regions were amplified by PCR from human genomic DNA and inserted into the KpnI and NheI sites of pGL-3 Basic (Promega, Madison, WI) to generate reporter plasmids.

    Article Title: Identification of Synaptotagmin 10 as Effector of NPAS4-Mediated Protection from Excitotoxic Neurodegeneration
    Article Snippet: The Syt10 promoter fragments were amplified by PCR from rat genomic DNA as template using primers with the corresponding restriction sites for subcloning into the luciferase reporter vector pGL-3 basic (Promega; pGL3–4713 FW: 5′-GCGACGCGTAGTGATTTGTAGAAAAGACCACATGA-3′, MluI; pGL3–1036 FW: 5′-GCGACGCGTGGGGTAACTTTACCATAACCTGG-3′, MluI; pGL3–306 FW: 5′-GCGACGCGTTCAGGAAGGTGTGTGGGAA-3′, MluI; pGL3–306 RV: 5′-GCGGTCGACCTTGGCTTCTGCTCGCAG-3′, SalI; RR-I FW: 5′-GCGGGATCCTAAAGGGACAAGCTTGGGCT-3′, XhoI; RR-I RV: 5′-GCGGGTCGACAAAATTCTTGATAAATACTTGCTGAA 3′, BglII; RR-5′ FW: 5′-GCGGGTACCTCTTCAGGCTCAAAATTAGTGG-3′, KpnI; RR-5′ RV: 5′-GCGGGTACCCAAACTTCAAAAACATCATGCTA-3′, KpnI; RR-3′ FW: 5′-GTCGACGTTCTTCTCGCAGAGCCTCA-3′, SalI; RR-3′ RV: 5′-GTCGACAAGTCGGGGAAAGGGATTATT-3′, SalI; RR-1/-3 FW: 5′-GCGGGTACCAGAAGTTTACCCAGCTCACCA-3′, KpnI; RR-1/-2 RV: 5′-GCGGGTACCACTCACTGTGGTAAGCATTCATT-3′, KpnI; RR-2 FW: 5′-GCGGGTACCCTGCCCAGACTGTAGATAAACG-3′, KpnI; RR-3 RV: 5′-GCGGGTACCTTATCTACAGTCTGGGCAGAACA-3′, KpnI; exon 1 FW: 5′-GCGCTCGAGTACCTGTGCCACCTCCG-3′, XhoI; exon 1 RV: 5′-GCGAAGCTTAGTTTCCGCAAGGAGGAC-3′, HindIII).

    Article Title: Constitutive Activation of Nrf2 in Mice Expands Enterogenesis in Small Intestine Through Negative Regulation of Math1
    Article Snippet: The target PCR products directly cloned between the KpnI and NcoI sites of pGL-3 basic (Promega).

    Article Title: PPARγ attenuates hypoxia-induced hypertrophic transcriptional pathways in the heart
    Article Snippet: This NFAT-Luc construct was designed using nine copies of an NFAT binding site from the IL-4 promoter (5’-TGGAAAATT-3’) positioned 5’ to a minimal promoter from the alpha-myosin heavy chain gene (–164 to +16) and inserted upstream of the luciferase reporter in pGL-3 Basic (Promega Corporation, Madison, WI, USA) to create NFAT-luc.

    Article Title: Identification of Synaptotagmin 10 as Effector of NPAS4-Mediated Protection from Excitotoxic Neurodegeneration
    Article Snippet: The Syt10 promoter fragments were amplified by PCR from rat genomic DNA as template using primers with the corresponding restriction sites for subcloning into the luciferase reporter vector pGL-3 basic (Promega; pGL3–4713 FW: 5 -GCGACGCGTAGT GATTTGTAGAAAAGACCACATGA-3 , MluI; pGL3–1036 FW: 5 -GCGACGCGTGGGGTAACTTTACCATAACCTGG-3 , MluI; pGL3–306 FW: 5 -GCGACGCGTTCAGGAAGGTGTGTGGGAA-3 , MluI; pGL3– 306 RV: 5 -GCGGTCGACCTTGGCTTCTGCTCGCAG-3 , SalI; RR-I FW: 5 -GCGGGATCCTAAAGGGACAAGCTTGGGCT-3 , XhoI; RR-I RV: 5 - GCGGGTCGACAAAATTCTTGATAAATACTTGCTGAA 3 , BglII; RR-5 FW: 5 -GCGGGTACCTCTTCAGGCTCAAAATTAGTGG-3 , KpnI; RR-5 RV: 5 -GCGGGTACCCAAACTTCAAAAACATCATGCTA-3 , KpnI; RR-3 FW: 5 -GTCGACGTTCTTCTCGCAGAGCCTCA-3 , SalI; RR-3 RV: 5 -GTCGACAAGTCGGGGAAAGGGATTATT-3 , SalI; RR1/-3 FW: 5 -GCGGGTACCAGAAGTTTACCCAGCTCACCA-3 , KpnI; RR-1/-2 RV: 5 -GCGGGTACCACTCACTGTGGTAAGCATTCATT-3 , KpnI; RR-2 FW: 5 -GCGGGTACCCTGCCCAGACTGTAGATAAACG-3 , KpnI; RR-3 RV: 5 -GCGGGTACCTTATCTACAGTCTGGGCAGAACA3 , KpnI; exon 1 FW: 5 -GCGCTCGAGTACCTGTGCCACCTCCG-3 , XhoI; exon 1 RV: 5 -GCGAAGCTTAGTTTCCGCAAGGAGGAC-3 , HindIII).

    Construct:

    Article Title: GATA2 Promotes Hematopoietic Development and Represses Cardiac Differentiation of Human Mesoderm
    Article Snippet: PITX2 and ISL1 promoter regions were amplified by PCR from human genomic DNA and inserted into the KpnI and NheI sites of pGL-3 Basic (Promega, Madison, WI) to generate reporter plasmids.

    Article Title: A circuit of COCH neurons encodes social-stress-induced anxiety via MTF1 activation of Cacna1h.
    Article Snippet: The sequence ofCacna1h 1020 promoter region was amplified by polymerase chain reaction(PCR) using mouse genomic DNA as a template with the primer set forward (MluI) (50-AT AGGTTGTCCAAGCAGTAAAGGG-30) and reverse (XhoI) (50-GGCTCACGTGTGCTCAGCC-30), and subcloned into the luciferase reporter vector pGL-3 basic (Promega).

    Article Title: Overexpression of homeodomain-interacting protein kinase 2 (HIPK2) attenuates sepsis-mediated liver injury by restoring autophagy
    Article Snippet: For further studies, primary hepatocytes were co-transfected with the internal control vector pmir-GLO (Promega) and pmir-GLO-HIPK2 3′-UTR using the Lipofectamine 2000 reagent (Thermo Fisher Scientific) for 20 h, followed by an incubation with the indicated chemicals for 10 h. Similarly, hepatocytes were co-transfected with the empty vector pGL-3 basic (Promega) and the pGL-3 vector containing the HIPK2 promoter using the Lipofectamine 2000 reagent for 20 h; the pRL vector was used as an internal control.

    Article Title: GATA2 Promotes Hematopoietic Development and Represses Cardiac Differentiation of Human Mesoderm
    Article Snippet: PITX2 and ISL1 promoter regions were amplified by PCR from human genomic DNA and inserted into the KpnI and NheI sites of pGL-3 Basic (Promega, Madison, WI) to generate reporter plasmids.

    Article Title: Identification of Synaptotagmin 10 as Effector of NPAS4-Mediated Protection from Excitotoxic Neurodegeneration
    Article Snippet: The Syt10 promoter fragments were amplified by PCR from rat genomic DNA as template using primers with the corresponding restriction sites for subcloning into the luciferase reporter vector pGL-3 basic (Promega; pGL3–4713 FW: 5′-GCGACGCGTAGTGATTTGTAGAAAAGACCACATGA-3′, MluI; pGL3–1036 FW: 5′-GCGACGCGTGGGGTAACTTTACCATAACCTGG-3′, MluI; pGL3–306 FW: 5′-GCGACGCGTTCAGGAAGGTGTGTGGGAA-3′, MluI; pGL3–306 RV: 5′-GCGGTCGACCTTGGCTTCTGCTCGCAG-3′, SalI; RR-I FW: 5′-GCGGGATCCTAAAGGGACAAGCTTGGGCT-3′, XhoI; RR-I RV: 5′-GCGGGTCGACAAAATTCTTGATAAATACTTGCTGAA 3′, BglII; RR-5′ FW: 5′-GCGGGTACCTCTTCAGGCTCAAAATTAGTGG-3′, KpnI; RR-5′ RV: 5′-GCGGGTACCCAAACTTCAAAAACATCATGCTA-3′, KpnI; RR-3′ FW: 5′-GTCGACGTTCTTCTCGCAGAGCCTCA-3′, SalI; RR-3′ RV: 5′-GTCGACAAGTCGGGGAAAGGGATTATT-3′, SalI; RR-1/-3 FW: 5′-GCGGGTACCAGAAGTTTACCCAGCTCACCA-3′, KpnI; RR-1/-2 RV: 5′-GCGGGTACCACTCACTGTGGTAAGCATTCATT-3′, KpnI; RR-2 FW: 5′-GCGGGTACCCTGCCCAGACTGTAGATAAACG-3′, KpnI; RR-3 RV: 5′-GCGGGTACCTTATCTACAGTCTGGGCAGAACA-3′, KpnI; exon 1 FW: 5′-GCGCTCGAGTACCTGTGCCACCTCCG-3′, XhoI; exon 1 RV: 5′-GCGAAGCTTAGTTTCCGCAAGGAGGAC-3′, HindIII).

    Article Title: Constitutive Activation of Nrf2 in Mice Expands Enterogenesis in Small Intestine Through Negative Regulation of Math1
    Article Snippet: The target PCR products directly cloned between the KpnI and NcoI sites of pGL-3 basic (Promega).

    Article Title: PPARγ attenuates hypoxia-induced hypertrophic transcriptional pathways in the heart
    Article Snippet: This NFAT-Luc construct was designed using nine copies of an NFAT binding site from the IL-4 promoter (5’-TGGAAAATT-3’) positioned 5’ to a minimal promoter from the alpha-myosin heavy chain gene (–164 to +16) and inserted upstream of the luciferase reporter in pGL-3 Basic (Promega Corporation, Madison, WI, USA) to create NFAT-luc.

    Article Title: Identification of Synaptotagmin 10 as Effector of NPAS4-Mediated Protection from Excitotoxic Neurodegeneration
    Article Snippet: The Syt10 promoter fragments were amplified by PCR from rat genomic DNA as template using primers with the corresponding restriction sites for subcloning into the luciferase reporter vector pGL-3 basic (Promega; pGL3–4713 FW: 5 -GCGACGCGTAGT GATTTGTAGAAAAGACCACATGA-3 , MluI; pGL3–1036 FW: 5 -GCGACGCGTGGGGTAACTTTACCATAACCTGG-3 , MluI; pGL3–306 FW: 5 -GCGACGCGTTCAGGAAGGTGTGTGGGAA-3 , MluI; pGL3– 306 RV: 5 -GCGGTCGACCTTGGCTTCTGCTCGCAG-3 , SalI; RR-I FW: 5 -GCGGGATCCTAAAGGGACAAGCTTGGGCT-3 , XhoI; RR-I RV: 5 - GCGGGTCGACAAAATTCTTGATAAATACTTGCTGAA 3 , BglII; RR-5 FW: 5 -GCGGGTACCTCTTCAGGCTCAAAATTAGTGG-3 , KpnI; RR-5 RV: 5 -GCGGGTACCCAAACTTCAAAAACATCATGCTA-3 , KpnI; RR-3 FW: 5 -GTCGACGTTCTTCTCGCAGAGCCTCA-3 , SalI; RR-3 RV: 5 -GTCGACAAGTCGGGGAAAGGGATTATT-3 , SalI; RR1/-3 FW: 5 -GCGGGTACCAGAAGTTTACCCAGCTCACCA-3 , KpnI; RR-1/-2 RV: 5 -GCGGGTACCACTCACTGTGGTAAGCATTCATT-3 , KpnI; RR-2 FW: 5 -GCGGGTACCCTGCCCAGACTGTAGATAAACG-3 , KpnI; RR-3 RV: 5 -GCGGGTACCTTATCTACAGTCTGGGCAGAACA3 , KpnI; exon 1 FW: 5 -GCGCTCGAGTACCTGTGCCACCTCCG-3 , XhoI; exon 1 RV: 5 -GCGAAGCTTAGTTTCCGCAAGGAGGAC-3 , HindIII).

    Binding Assay:

    Article Title: GATA2 Promotes Hematopoietic Development and Represses Cardiac Differentiation of Human Mesoderm
    Article Snippet: PITX2 and ISL1 promoter regions were amplified by PCR from human genomic DNA and inserted into the KpnI and NheI sites of pGL-3 Basic (Promega, Madison, WI) to generate reporter plasmids.

    Article Title: A circuit of COCH neurons encodes social-stress-induced anxiety via MTF1 activation of Cacna1h.
    Article Snippet: The sequence ofCacna1h 1020 promoter region was amplified by polymerase chain reaction(PCR) using mouse genomic DNA as a template with the primer set forward (MluI) (50-AT AGGTTGTCCAAGCAGTAAAGGG-30) and reverse (XhoI) (50-GGCTCACGTGTGCTCAGCC-30), and subcloned into the luciferase reporter vector pGL-3 basic (Promega).

    Article Title: Overexpression of homeodomain-interacting protein kinase 2 (HIPK2) attenuates sepsis-mediated liver injury by restoring autophagy
    Article Snippet: For further studies, primary hepatocytes were co-transfected with the internal control vector pmir-GLO (Promega) and pmir-GLO-HIPK2 3′-UTR using the Lipofectamine 2000 reagent (Thermo Fisher Scientific) for 20 h, followed by an incubation with the indicated chemicals for 10 h. Similarly, hepatocytes were co-transfected with the empty vector pGL-3 basic (Promega) and the pGL-3 vector containing the HIPK2 promoter using the Lipofectamine 2000 reagent for 20 h; the pRL vector was used as an internal control.

    Article Title: GATA2 Promotes Hematopoietic Development and Represses Cardiac Differentiation of Human Mesoderm
    Article Snippet: PITX2 and ISL1 promoter regions were amplified by PCR from human genomic DNA and inserted into the KpnI and NheI sites of pGL-3 Basic (Promega, Madison, WI) to generate reporter plasmids.

    Article Title: Identification of Synaptotagmin 10 as Effector of NPAS4-Mediated Protection from Excitotoxic Neurodegeneration
    Article Snippet: The Syt10 promoter fragments were amplified by PCR from rat genomic DNA as template using primers with the corresponding restriction sites for subcloning into the luciferase reporter vector pGL-3 basic (Promega; pGL3–4713 FW: 5′-GCGACGCGTAGTGATTTGTAGAAAAGACCACATGA-3′, MluI; pGL3–1036 FW: 5′-GCGACGCGTGGGGTAACTTTACCATAACCTGG-3′, MluI; pGL3–306 FW: 5′-GCGACGCGTTCAGGAAGGTGTGTGGGAA-3′, MluI; pGL3–306 RV: 5′-GCGGTCGACCTTGGCTTCTGCTCGCAG-3′, SalI; RR-I FW: 5′-GCGGGATCCTAAAGGGACAAGCTTGGGCT-3′, XhoI; RR-I RV: 5′-GCGGGTCGACAAAATTCTTGATAAATACTTGCTGAA 3′, BglII; RR-5′ FW: 5′-GCGGGTACCTCTTCAGGCTCAAAATTAGTGG-3′, KpnI; RR-5′ RV: 5′-GCGGGTACCCAAACTTCAAAAACATCATGCTA-3′, KpnI; RR-3′ FW: 5′-GTCGACGTTCTTCTCGCAGAGCCTCA-3′, SalI; RR-3′ RV: 5′-GTCGACAAGTCGGGGAAAGGGATTATT-3′, SalI; RR-1/-3 FW: 5′-GCGGGTACCAGAAGTTTACCCAGCTCACCA-3′, KpnI; RR-1/-2 RV: 5′-GCGGGTACCACTCACTGTGGTAAGCATTCATT-3′, KpnI; RR-2 FW: 5′-GCGGGTACCCTGCCCAGACTGTAGATAAACG-3′, KpnI; RR-3 RV: 5′-GCGGGTACCTTATCTACAGTCTGGGCAGAACA-3′, KpnI; exon 1 FW: 5′-GCGCTCGAGTACCTGTGCCACCTCCG-3′, XhoI; exon 1 RV: 5′-GCGAAGCTTAGTTTCCGCAAGGAGGAC-3′, HindIII).

    Article Title: Constitutive Activation of Nrf2 in Mice Expands Enterogenesis in Small Intestine Through Negative Regulation of Math1
    Article Snippet: The target PCR products directly cloned between the KpnI and NcoI sites of pGL-3 basic (Promega).

    Article Title: PPARγ attenuates hypoxia-induced hypertrophic transcriptional pathways in the heart
    Article Snippet: This NFAT-Luc construct was designed using nine copies of an NFAT binding site from the IL-4 promoter (5’-TGGAAAATT-3’) positioned 5’ to a minimal promoter from the alpha-myosin heavy chain gene (–164 to +16) and inserted upstream of the luciferase reporter in pGL-3 Basic (Promega Corporation, Madison, WI, USA) to create NFAT-luc.

    Article Title: Identification of Synaptotagmin 10 as Effector of NPAS4-Mediated Protection from Excitotoxic Neurodegeneration
    Article Snippet: The Syt10 promoter fragments were amplified by PCR from rat genomic DNA as template using primers with the corresponding restriction sites for subcloning into the luciferase reporter vector pGL-3 basic (Promega; pGL3–4713 FW: 5 -GCGACGCGTAGT GATTTGTAGAAAAGACCACATGA-3 , MluI; pGL3–1036 FW: 5 -GCGACGCGTGGGGTAACTTTACCATAACCTGG-3 , MluI; pGL3–306 FW: 5 -GCGACGCGTTCAGGAAGGTGTGTGGGAA-3 , MluI; pGL3– 306 RV: 5 -GCGGTCGACCTTGGCTTCTGCTCGCAG-3 , SalI; RR-I FW: 5 -GCGGGATCCTAAAGGGACAAGCTTGGGCT-3 , XhoI; RR-I RV: 5 - GCGGGTCGACAAAATTCTTGATAAATACTTGCTGAA 3 , BglII; RR-5 FW: 5 -GCGGGTACCTCTTCAGGCTCAAAATTAGTGG-3 , KpnI; RR-5 RV: 5 -GCGGGTACCCAAACTTCAAAAACATCATGCTA-3 , KpnI; RR-3 FW: 5 -GTCGACGTTCTTCTCGCAGAGCCTCA-3 , SalI; RR-3 RV: 5 -GTCGACAAGTCGGGGAAAGGGATTATT-3 , SalI; RR1/-3 FW: 5 -GCGGGTACCAGAAGTTTACCCAGCTCACCA-3 , KpnI; RR-1/-2 RV: 5 -GCGGGTACCACTCACTGTGGTAAGCATTCATT-3 , KpnI; RR-2 FW: 5 -GCGGGTACCCTGCCCAGACTGTAGATAAACG-3 , KpnI; RR-3 RV: 5 -GCGGGTACCTTATCTACAGTCTGGGCAGAACA3 , KpnI; exon 1 FW: 5 -GCGCTCGAGTACCTGTGCCACCTCCG-3 , XhoI; exon 1 RV: 5 -GCGAAGCTTAGTTTCCGCAAGGAGGAC-3 , HindIII).

    Subcloning:

    Article Title: GATA2 Promotes Hematopoietic Development and Represses Cardiac Differentiation of Human Mesoderm
    Article Snippet: PITX2 and ISL1 promoter regions were amplified by PCR from human genomic DNA and inserted into the KpnI and NheI sites of pGL-3 Basic (Promega, Madison, WI) to generate reporter plasmids.

    Article Title: A circuit of COCH neurons encodes social-stress-induced anxiety via MTF1 activation of Cacna1h.
    Article Snippet: The sequence ofCacna1h 1020 promoter region was amplified by polymerase chain reaction(PCR) using mouse genomic DNA as a template with the primer set forward (MluI) (50-AT AGGTTGTCCAAGCAGTAAAGGG-30) and reverse (XhoI) (50-GGCTCACGTGTGCTCAGCC-30), and subcloned into the luciferase reporter vector pGL-3 basic (Promega).

    Article Title: Overexpression of homeodomain-interacting protein kinase 2 (HIPK2) attenuates sepsis-mediated liver injury by restoring autophagy
    Article Snippet: For further studies, primary hepatocytes were co-transfected with the internal control vector pmir-GLO (Promega) and pmir-GLO-HIPK2 3′-UTR using the Lipofectamine 2000 reagent (Thermo Fisher Scientific) for 20 h, followed by an incubation with the indicated chemicals for 10 h. Similarly, hepatocytes were co-transfected with the empty vector pGL-3 basic (Promega) and the pGL-3 vector containing the HIPK2 promoter using the Lipofectamine 2000 reagent for 20 h; the pRL vector was used as an internal control.

    Article Title: GATA2 Promotes Hematopoietic Development and Represses Cardiac Differentiation of Human Mesoderm
    Article Snippet: PITX2 and ISL1 promoter regions were amplified by PCR from human genomic DNA and inserted into the KpnI and NheI sites of pGL-3 Basic (Promega, Madison, WI) to generate reporter plasmids.

    Article Title: Identification of Synaptotagmin 10 as Effector of NPAS4-Mediated Protection from Excitotoxic Neurodegeneration
    Article Snippet: The Syt10 promoter fragments were amplified by PCR from rat genomic DNA as template using primers with the corresponding restriction sites for subcloning into the luciferase reporter vector pGL-3 basic (Promega; pGL3–4713 FW: 5′-GCGACGCGTAGTGATTTGTAGAAAAGACCACATGA-3′, MluI; pGL3–1036 FW: 5′-GCGACGCGTGGGGTAACTTTACCATAACCTGG-3′, MluI; pGL3–306 FW: 5′-GCGACGCGTTCAGGAAGGTGTGTGGGAA-3′, MluI; pGL3–306 RV: 5′-GCGGTCGACCTTGGCTTCTGCTCGCAG-3′, SalI; RR-I FW: 5′-GCGGGATCCTAAAGGGACAAGCTTGGGCT-3′, XhoI; RR-I RV: 5′-GCGGGTCGACAAAATTCTTGATAAATACTTGCTGAA 3′, BglII; RR-5′ FW: 5′-GCGGGTACCTCTTCAGGCTCAAAATTAGTGG-3′, KpnI; RR-5′ RV: 5′-GCGGGTACCCAAACTTCAAAAACATCATGCTA-3′, KpnI; RR-3′ FW: 5′-GTCGACGTTCTTCTCGCAGAGCCTCA-3′, SalI; RR-3′ RV: 5′-GTCGACAAGTCGGGGAAAGGGATTATT-3′, SalI; RR-1/-3 FW: 5′-GCGGGTACCAGAAGTTTACCCAGCTCACCA-3′, KpnI; RR-1/-2 RV: 5′-GCGGGTACCACTCACTGTGGTAAGCATTCATT-3′, KpnI; RR-2 FW: 5′-GCGGGTACCCTGCCCAGACTGTAGATAAACG-3′, KpnI; RR-3 RV: 5′-GCGGGTACCTTATCTACAGTCTGGGCAGAACA-3′, KpnI; exon 1 FW: 5′-GCGCTCGAGTACCTGTGCCACCTCCG-3′, XhoI; exon 1 RV: 5′-GCGAAGCTTAGTTTCCGCAAGGAGGAC-3′, HindIII).

    Article Title: Constitutive Activation of Nrf2 in Mice Expands Enterogenesis in Small Intestine Through Negative Regulation of Math1
    Article Snippet: The target PCR products directly cloned between the KpnI and NcoI sites of pGL-3 basic (Promega).

    Article Title: PPARγ attenuates hypoxia-induced hypertrophic transcriptional pathways in the heart
    Article Snippet: This NFAT-Luc construct was designed using nine copies of an NFAT binding site from the IL-4 promoter (5’-TGGAAAATT-3’) positioned 5’ to a minimal promoter from the alpha-myosin heavy chain gene (–164 to +16) and inserted upstream of the luciferase reporter in pGL-3 Basic (Promega Corporation, Madison, WI, USA) to create NFAT-luc.

    Article Title: Identification of Synaptotagmin 10 as Effector of NPAS4-Mediated Protection from Excitotoxic Neurodegeneration
    Article Snippet: The Syt10 promoter fragments were amplified by PCR from rat genomic DNA as template using primers with the corresponding restriction sites for subcloning into the luciferase reporter vector pGL-3 basic (Promega; pGL3–4713 FW: 5 -GCGACGCGTAGT GATTTGTAGAAAAGACCACATGA-3 , MluI; pGL3–1036 FW: 5 -GCGACGCGTGGGGTAACTTTACCATAACCTGG-3 , MluI; pGL3–306 FW: 5 -GCGACGCGTTCAGGAAGGTGTGTGGGAA-3 , MluI; pGL3– 306 RV: 5 -GCGGTCGACCTTGGCTTCTGCTCGCAG-3 , SalI; RR-I FW: 5 -GCGGGATCCTAAAGGGACAAGCTTGGGCT-3 , XhoI; RR-I RV: 5 - GCGGGTCGACAAAATTCTTGATAAATACTTGCTGAA 3 , BglII; RR-5 FW: 5 -GCGGGTACCTCTTCAGGCTCAAAATTAGTGG-3 , KpnI; RR-5 RV: 5 -GCGGGTACCCAAACTTCAAAAACATCATGCTA-3 , KpnI; RR-3 FW: 5 -GTCGACGTTCTTCTCGCAGAGCCTCA-3 , SalI; RR-3 RV: 5 -GTCGACAAGTCGGGGAAAGGGATTATT-3 , SalI; RR1/-3 FW: 5 -GCGGGTACCAGAAGTTTACCCAGCTCACCA-3 , KpnI; RR-1/-2 RV: 5 -GCGGGTACCACTCACTGTGGTAAGCATTCATT-3 , KpnI; RR-2 FW: 5 -GCGGGTACCCTGCCCAGACTGTAGATAAACG-3 , KpnI; RR-3 RV: 5 -GCGGGTACCTTATCTACAGTCTGGGCAGAACA3 , KpnI; exon 1 FW: 5 -GCGCTCGAGTACCTGTGCCACCTCCG-3 , XhoI; exon 1 RV: 5 -GCGAAGCTTAGTTTCCGCAAGGAGGAC-3 , HindIII).



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    The (−2436/+149) region of the PAK3 promoter, containing several putative cJun/AP- binding sites, was cloned into the pGL 3 -Basic reporter vector. A: Luciferase promoter reporter assays performed in control cells, Rat1a-GFP, and Rat1a-J4 cells transiently transfected with empty vector, pGL 3 -Basic, and PAK3 promoter construct containing vector, pGL 3 -Basic-pPAK3 (−2436/+149). Cells were grown in the absence and presence of doxycycline. B: Luciferase promoter reporter assays performed in the parental rat fibroblast cell line, Rat1a, transfected with either pCMV-cJun or empty pCMV vector with the PAK3 promoter construct containing vector, pGL 3 -Basic-pPAK3 (−2436/+149). A plasmid containing four AP-1 binding sites, 4 X AP-1-Luc, was used as a control showing cJun activation. C: Luciferase reporter promoter assays for deletion constructs of the PAK3 promoter region (−2436/+149), (−2329/+149), (−684/+149) and (−179/+149) as well as a (−179/+149) with a mutated cJun/AP-1 binding site at position (+52/+60), in the presence and absence of doxycycline-induced cJun/AP-1 expression. D: Luciferase reporter assays, using the (−179/+149) PAK3 promoter construct, showing the effect of transient cJun inhibition on PAK3 promoter activity in the absence and presence of doxycycline induced cJun/AP-1 over-expression. Results show the mean±S.E. of experiments performed in triplicate and repeated at least three times. (*p≤0.05 and **p≤0.01).

    Journal: PLoS ONE

    Article Title: p21-Activated Kinase 3 (PAK3) Is an AP-1 Regulated Gene Contributing to Actin Organisation and Migration of Transformed Fibroblasts

    doi: 10.1371/journal.pone.0066892

    Figure Lengend Snippet: The (−2436/+149) region of the PAK3 promoter, containing several putative cJun/AP- binding sites, was cloned into the pGL 3 -Basic reporter vector. A: Luciferase promoter reporter assays performed in control cells, Rat1a-GFP, and Rat1a-J4 cells transiently transfected with empty vector, pGL 3 -Basic, and PAK3 promoter construct containing vector, pGL 3 -Basic-pPAK3 (−2436/+149). Cells were grown in the absence and presence of doxycycline. B: Luciferase promoter reporter assays performed in the parental rat fibroblast cell line, Rat1a, transfected with either pCMV-cJun or empty pCMV vector with the PAK3 promoter construct containing vector, pGL 3 -Basic-pPAK3 (−2436/+149). A plasmid containing four AP-1 binding sites, 4 X AP-1-Luc, was used as a control showing cJun activation. C: Luciferase reporter promoter assays for deletion constructs of the PAK3 promoter region (−2436/+149), (−2329/+149), (−684/+149) and (−179/+149) as well as a (−179/+149) with a mutated cJun/AP-1 binding site at position (+52/+60), in the presence and absence of doxycycline-induced cJun/AP-1 expression. D: Luciferase reporter assays, using the (−179/+149) PAK3 promoter construct, showing the effect of transient cJun inhibition on PAK3 promoter activity in the absence and presence of doxycycline induced cJun/AP-1 over-expression. Results show the mean±S.E. of experiments performed in triplicate and repeated at least three times. (*p≤0.05 and **p≤0.01).

    Article Snippet: PCR using high fidelity Expand Plus DNA Polymerase (Roche) was performed on rat genomic DNA, the products were sub-cloned into the pGEM-T Easy vector (Promega) and excised for cloning into the luciferase reported vector, pGL 3 -Basic (Promega) using Mlu1 and Xho1 restriction enzymes.

    Techniques: Binding Assay, Clone Assay, Plasmid Preparation, Luciferase, Transfection, Construct, Activation Assay, Expressing, Inhibition, Activity Assay, Over Expression