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pgl3basic vectors  (Promega)

 
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    Promega pgl3basic vectors
    Pgl3basic Vectors, 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/pgl3basic+vectors/pgl3+basic/pm40253379-413-25-27
    Average 90 stars, based on 1 article reviews
    pgl3basic vectors - by Bioz Stars, 2026-10
    90/100 stars

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    Related Articles

    other:

    Article Title: Role of Hypoxia-inducible Factor-1 in Transcriptional Activation of Ceruloplasmin by Iron Deficiency
    Article Snippet: The constructs were ligated into SacI and XhoI sites of pGL3basic or pGL3prom vectors (Promega).

    Article Title: Functional genetic variants of the IFN-λ3 (IL28B) gene and transcription factor interactions on its promoter
    Article Snippet: We amplified the 1.4 kb IFN-λ3 promoter and 5’UTR DNA from five HCV-infected patients from our cohort (described in Ref. [ ]) and cloned it in pGL3basic (Promega) vectors.

    Clone Assay:

    Article Title: Vitellogenin receptor mediates heat adaptability of oocyte development in mud crabs and zebrafish.
    Article Snippet: .. The enhancer sequences of VtgR from mud crabs, which show differential H3K27ac signals in crabs maintained at 25 °C and 33 °C, were cloned into pGL3Basic vectors (Promega, USA) along with the promoter sequence of VtgR using the primers listed in Supplementary Table 15. .. After DNA quantification and qualification, all libraries were sequenced with 2× 150bp chemistry on the Illumina Nova-seq 6000 platform (Illumina, USA).

    Article Title: Identification of a gene variant in the master regulator of lipid metabolism SREBP-1 in a family with a novel form of severe combined hypolipidemia.
    Article Snippet: Objective: Alterations of lipid metabolism play a pivotal role in the development of atherosclerosis and its complications, today’s major mortality risks.. The predominant regulators controlling cholesteroland fatty acids synthesis in liver are the sterol regulatory element-binding proteins (SREBPs), a family of transcription factors that were formerly identified as cholesterol sensor for LDLR gene expression.. Variation of gene structure in these genes might therefore indicate a predisposition to develop complications like myocardial infarction and stroke.

    Article Title: miR-363-5p protects from neuropathic pain in chronic constriction injury (CCI) rat models and regulates Schwann cell injury via negatively modulating SERPING1.
    Article Snippet: Objectives: Due to the complex and unclear pathogenesis of neuropathic pain, there is a lack of effective therapeutic strategy. miR-363-5p was considered of great potential in mediating the development of neuropathic pain, which has not been confirmed with direct evidence.. This study evaluated the role of miR-363-5p in neuropathic pain with animal and cell models, aiming to reveal the potential of miR-363-5p in target therapy of neuropathic pain.. Methods: Chronic constriction injury (CCI) rat models were established as the neuropathic pain model.

    Sequencing:

    Article Title: Vitellogenin receptor mediates heat adaptability of oocyte development in mud crabs and zebrafish.
    Article Snippet: .. The enhancer sequences of VtgR from mud crabs, which show differential H3K27ac signals in crabs maintained at 25 °C and 33 °C, were cloned into pGL3Basic vectors (Promega, USA) along with the promoter sequence of VtgR using the primers listed in Supplementary Table 15. .. After DNA quantification and qualification, all libraries were sequenced with 2× 150bp chemistry on the Illumina Nova-seq 6000 platform (Illumina, USA).

    Polymerase Chain Reaction:

    Article Title: Identification of a gene variant in the master regulator of lipid metabolism SREBP-1 in a family with a novel form of severe combined hypolipidemia.
    Article Snippet: Objective: Alterations of lipid metabolism play a pivotal role in the development of atherosclerosis and its complications, today’s major mortality risks.. The predominant regulators controlling cholesteroland fatty acids synthesis in liver are the sterol regulatory element-binding proteins (SREBPs), a family of transcription factors that were formerly identified as cholesterol sensor for LDLR gene expression.. Variation of gene structure in these genes might therefore indicate a predisposition to develop complications like myocardial infarction and stroke.

    Article Title: Genotypic Effect of ABCG1 gene promoter -257T>G polymorphism on coronary artery disease severity in Japanese men.
    Article Snippet: .. Human ABCG1 promoter constructs-Firefly luciferase reporter constructs were generated by PCR amplification of a 1141-bp fragment of human ABCG1 promoter (-1104/+37) and then subcloned with pGL3Basic vectors (Promega, Madison, WI), as described previously22). .. Site-directed mutagenesis-Mutant plasmids were obtained for human ABCG1 promoter using the QuickChange® Site-Directed Mutagenesis Kit (Stratagene, La Jolla, CA) and each of the following double-stranded oligonucleotides.

    Amplification:

    Article Title: Identification of a gene variant in the master regulator of lipid metabolism SREBP-1 in a family with a novel form of severe combined hypolipidemia.
    Article Snippet: Objective: Alterations of lipid metabolism play a pivotal role in the development of atherosclerosis and its complications, today’s major mortality risks.. The predominant regulators controlling cholesteroland fatty acids synthesis in liver are the sterol regulatory element-binding proteins (SREBPs), a family of transcription factors that were formerly identified as cholesterol sensor for LDLR gene expression.. Variation of gene structure in these genes might therefore indicate a predisposition to develop complications like myocardial infarction and stroke.

    Article Title: Genotypic Effect of ABCG1 gene promoter -257T>G polymorphism on coronary artery disease severity in Japanese men.
    Article Snippet: .. Human ABCG1 promoter constructs-Firefly luciferase reporter constructs were generated by PCR amplification of a 1141-bp fragment of human ABCG1 promoter (-1104/+37) and then subcloned with pGL3Basic vectors (Promega, Madison, WI), as described previously22). .. Site-directed mutagenesis-Mutant plasmids were obtained for human ABCG1 promoter using the QuickChange® Site-Directed Mutagenesis Kit (Stratagene, La Jolla, CA) and each of the following double-stranded oligonucleotides.

    Mutagenesis:

    Article Title: miR-363-5p protects from neuropathic pain in chronic constriction injury (CCI) rat models and regulates Schwann cell injury via negatively modulating SERPING1.
    Article Snippet: Objectives: Due to the complex and unclear pathogenesis of neuropathic pain, there is a lack of effective therapeutic strategy. miR-363-5p was considered of great potential in mediating the development of neuropathic pain, which has not been confirmed with direct evidence.. This study evaluated the role of miR-363-5p in neuropathic pain with animal and cell models, aiming to reveal the potential of miR-363-5p in target therapy of neuropathic pain.. Methods: Chronic constriction injury (CCI) rat models were established as the neuropathic pain model.

    Article Title: STAT3-induced ZBED3-AS1 promotes the malignant phenotypes of melanoma cells by activating PI3K/AKT signaling pathway.
    Article Snippet: .. The ZBED3-AS1 promoter sequences with wild-type (WT) or mutant (MUT) STAT3 binding sites were inserted to pGL3basic vectors (Promega, Madison, WI, USA). ..

    Construct:

    Article Title: Genotypic Effect of ABCG1 gene promoter -257T>G polymorphism on coronary artery disease severity in Japanese men.
    Article Snippet: .. Human ABCG1 promoter constructs-Firefly luciferase reporter constructs were generated by PCR amplification of a 1141-bp fragment of human ABCG1 promoter (-1104/+37) and then subcloned with pGL3Basic vectors (Promega, Madison, WI), as described previously22). .. Site-directed mutagenesis-Mutant plasmids were obtained for human ABCG1 promoter using the QuickChange® Site-Directed Mutagenesis Kit (Stratagene, La Jolla, CA) and each of the following double-stranded oligonucleotides.

    Article Title: Nrf2 pathway in vegetable oil-induced inflammation of large yellow croaker (Larimichthys crocea).
    Article Snippet: This study was conducted to investigate the effects and regulation of dietary vegetable oil (VO, enriched with α-linolenic acid [ALA] and linoleic acid [LNA]) on the nuclear factor erythroid 2-related factor 2 (Nrf2) and nuclear factor-κB (NF-κB) pathways in large yellow croaker.. In vivo study showed that the VO diet significantly decreased the activity of antioxidant enzymes and antioxidant enzyme-related mRNA expression in the liver tissue, in comparison with the fish oil (FO) diet (P < 0.05).. The suppression of antioxidant capacity might be due to the decrease of nuclear Nrf2 protein translocation, Nrf2 binding to antioxidant response element (ARE) sequences, and subsequently, antioxidant genes transcription as electrophoretic mobility shift assay (EMSA) and luciferase assay showed.

    Luciferase:

    Article Title: Genotypic Effect of ABCG1 gene promoter -257T>G polymorphism on coronary artery disease severity in Japanese men.
    Article Snippet: .. Human ABCG1 promoter constructs-Firefly luciferase reporter constructs were generated by PCR amplification of a 1141-bp fragment of human ABCG1 promoter (-1104/+37) and then subcloned with pGL3Basic vectors (Promega, Madison, WI), as described previously22). .. Site-directed mutagenesis-Mutant plasmids were obtained for human ABCG1 promoter using the QuickChange® Site-Directed Mutagenesis Kit (Stratagene, La Jolla, CA) and each of the following double-stranded oligonucleotides.

    Generated:

    Article Title: Genotypic Effect of ABCG1 gene promoter -257T>G polymorphism on coronary artery disease severity in Japanese men.
    Article Snippet: .. Human ABCG1 promoter constructs-Firefly luciferase reporter constructs were generated by PCR amplification of a 1141-bp fragment of human ABCG1 promoter (-1104/+37) and then subcloned with pGL3Basic vectors (Promega, Madison, WI), as described previously22). .. Site-directed mutagenesis-Mutant plasmids were obtained for human ABCG1 promoter using the QuickChange® Site-Directed Mutagenesis Kit (Stratagene, La Jolla, CA) and each of the following double-stranded oligonucleotides.

    Binding Assay:

    Article Title: STAT3-induced ZBED3-AS1 promotes the malignant phenotypes of melanoma cells by activating PI3K/AKT signaling pathway.
    Article Snippet: .. The ZBED3-AS1 promoter sequences with wild-type (WT) or mutant (MUT) STAT3 binding sites were inserted to pGL3basic vectors (Promega, Madison, WI, USA). ..



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    pgl3basic luciferase reporter vector  (Promega)
    90
    Promega pgl3basic luciferase reporter vector
    Piezo1 suppresses Lcn2 expression and secretion via the inhibition of Ccl2-evoked NF-κB activation. BMMSCs were isolated from femurs and tibias of 10-week-old male PDGFRα-Piezo1 KO mice and WT controls. a Gene expression of Ccr2 in BMMSCs, as determined by real-time PCR. b–g Recombinant mouse Ccl2 protein (rmCcl2, 100 ng/mL) or the CCR2 antagonist INCB3344 (10 nM) was added to the adipogenic and osteogenic induction medium during BMMSC differentiation. The expression levels of osteogenic genes ( b–d ) and adipogenic genes ( e–g ) were determined via real-time qPCR. h–n WT and KO BMMSCs were treated with rmCcl2 (100 ng/mL) or the CCR2 antagonist INCB3344 (10 nM) for 24 h. h Sonicated BMMSCs were subjected to chromatin immunoprecipitation (ChIP) using an anti-p65 antibody. The ChIP DNA samples were then subjected to qPCR amplification using specific primers against the promoter region of Lcn2, as shown in supplementary Fig. . Representative gel images of end-point ChIP-qPCR products are shown. i Quantitative results are shown as the percentage of input DNA: % of Input = 2^(CT Input −CT IP )/dilution factor × 100%. j <t>Luciferase</t> reporter assay was performed in isolated WT and KO BMMSCs transfected with pGL3 constructs containing the mouse wild-type Lcn2 promoter (Lcn2-WT) or the mutated Lcn2 promoter with mutations in the NF-κB binding motif (Lcn2-Mut, sequence in supplementary Fig. ). The firefly luciferase activity was measured and normalized to <t>Renilla</t> luciferase activity. k Schematic diagram illustrating the actions of rmCcl2, the CCR2 antagonist, and the NF-κB-specific inhibitor QNZ on NF-κB translocation and Lcn2 expression; created with BioRender ( https://BioRender.com ). l ELISA analysis of total p65 in the nucleus and cytoplasm of BMMSCs. m Representative immunofluorescence images of intracellular p65 localization. Nuclei were stained with DAPI (blue). Scale bar, 50 μm. n P65 nuclear translocation was quantified by ImageJ software via Pearson’s correlation coefficient (PCC) between the p65 immunofluorescence signal and DAPI staining across segmented nuclear regions. o , p WT and KO BMMSCs were treated with the NF-κB-specific inhibitor QNZ (10 nM) for another 2 h after 24-h treatment of rmCcl2 (100 ng/mL) and INCB3344 (10 nM). o The mRNA level of Lcn2 . p Lcn2 levels in the culture medium were determined by ELISA. q Illustration of how the AP-1 inhibitor modulates Ccl2 gene expression; created with BioRender ( https://BioRender.com ). r–u WT and KO BMMSCs were treated with the AP-1-specific inhibitor T-5224 (10 μM) for 24 h. r Sonicated BMMSCs were subjected to ChIP assay using an anti-c-Jun antibody. Representative gel images of end-point ChIP-qPCR products are shown. s Quantitative results are shown as the percentage of input DNA. t The mRNA level of Ccl2 . u Ccl2 levels in the culture medium were determined by ELISA. v Luciferase reporter assay was performed in isolated WT and KO BMMSCs transfected with pGL3 constructs containing the mouse wild-type Ccl2 promoter (Ccl2-WT) or the mutated Ccl2 promoter with mutations in the c-Jun binding motif (Ccl2-Mut, sequence in supplementary Fig. ). The firefly luciferase activity was measured and normalized to Renilla luciferase activity. n = 5 for each group. The data are presented as the means ± SEMs; * p < 0.05, ** p < 0.01, *** p < 0.001. See also supplementary Figs. –
    Pgl3basic Luciferase Reporter Vector, 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
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    Piezo1 suppresses Lcn2 expression and secretion via the inhibition of Ccl2-evoked NF-κB activation. BMMSCs were isolated from femurs and tibias of 10-week-old male PDGFRα-Piezo1 KO mice and WT controls. a Gene expression of Ccr2 in BMMSCs, as determined by real-time PCR. b–g Recombinant mouse Ccl2 protein (rmCcl2, 100 ng/mL) or the CCR2 antagonist INCB3344 (10 nM) was added to the adipogenic and osteogenic induction medium during BMMSC differentiation. The expression levels of osteogenic genes ( b–d ) and adipogenic genes ( e–g ) were determined via real-time qPCR. h–n WT and KO BMMSCs were treated with rmCcl2 (100 ng/mL) or the CCR2 antagonist INCB3344 (10 nM) for 24 h. h Sonicated BMMSCs were subjected to chromatin immunoprecipitation (ChIP) using an anti-p65 antibody. The ChIP DNA samples were then subjected to qPCR amplification using specific primers against the promoter region of Lcn2, as shown in supplementary Fig. . Representative gel images of end-point ChIP-qPCR products are shown. i Quantitative results are shown as the percentage of input DNA: % of Input = 2^(CT Input −CT IP )/dilution factor × 100%. j Luciferase reporter assay was performed in isolated WT and KO BMMSCs transfected with pGL3 constructs containing the mouse wild-type Lcn2 promoter (Lcn2-WT) or the mutated Lcn2 promoter with mutations in the NF-κB binding motif (Lcn2-Mut, sequence in supplementary Fig. ). The firefly luciferase activity was measured and normalized to Renilla luciferase activity. k Schematic diagram illustrating the actions of rmCcl2, the CCR2 antagonist, and the NF-κB-specific inhibitor QNZ on NF-κB translocation and Lcn2 expression; created with BioRender ( https://BioRender.com ). l ELISA analysis of total p65 in the nucleus and cytoplasm of BMMSCs. m Representative immunofluorescence images of intracellular p65 localization. Nuclei were stained with DAPI (blue). Scale bar, 50 μm. n P65 nuclear translocation was quantified by ImageJ software via Pearson’s correlation coefficient (PCC) between the p65 immunofluorescence signal and DAPI staining across segmented nuclear regions. o , p WT and KO BMMSCs were treated with the NF-κB-specific inhibitor QNZ (10 nM) for another 2 h after 24-h treatment of rmCcl2 (100 ng/mL) and INCB3344 (10 nM). o The mRNA level of Lcn2 . p Lcn2 levels in the culture medium were determined by ELISA. q Illustration of how the AP-1 inhibitor modulates Ccl2 gene expression; created with BioRender ( https://BioRender.com ). r–u WT and KO BMMSCs were treated with the AP-1-specific inhibitor T-5224 (10 μM) for 24 h. r Sonicated BMMSCs were subjected to ChIP assay using an anti-c-Jun antibody. Representative gel images of end-point ChIP-qPCR products are shown. s Quantitative results are shown as the percentage of input DNA. t The mRNA level of Ccl2 . u Ccl2 levels in the culture medium were determined by ELISA. v Luciferase reporter assay was performed in isolated WT and KO BMMSCs transfected with pGL3 constructs containing the mouse wild-type Ccl2 promoter (Ccl2-WT) or the mutated Ccl2 promoter with mutations in the c-Jun binding motif (Ccl2-Mut, sequence in supplementary Fig. ). The firefly luciferase activity was measured and normalized to Renilla luciferase activity. n = 5 for each group. The data are presented as the means ± SEMs; * p < 0.05, ** p < 0.01, *** p < 0.001. See also supplementary Figs. –

    Journal: Signal Transduction and Targeted Therapy

    Article Title: Piezo1 activation suppresses bone marrow adipogenesis to prevent osteoporosis by inhibiting a mechanoinflammatory autocrine loop

    doi: 10.1038/s41392-025-02455-w

    Figure Lengend Snippet: Piezo1 suppresses Lcn2 expression and secretion via the inhibition of Ccl2-evoked NF-κB activation. BMMSCs were isolated from femurs and tibias of 10-week-old male PDGFRα-Piezo1 KO mice and WT controls. a Gene expression of Ccr2 in BMMSCs, as determined by real-time PCR. b–g Recombinant mouse Ccl2 protein (rmCcl2, 100 ng/mL) or the CCR2 antagonist INCB3344 (10 nM) was added to the adipogenic and osteogenic induction medium during BMMSC differentiation. The expression levels of osteogenic genes ( b–d ) and adipogenic genes ( e–g ) were determined via real-time qPCR. h–n WT and KO BMMSCs were treated with rmCcl2 (100 ng/mL) or the CCR2 antagonist INCB3344 (10 nM) for 24 h. h Sonicated BMMSCs were subjected to chromatin immunoprecipitation (ChIP) using an anti-p65 antibody. The ChIP DNA samples were then subjected to qPCR amplification using specific primers against the promoter region of Lcn2, as shown in supplementary Fig. . Representative gel images of end-point ChIP-qPCR products are shown. i Quantitative results are shown as the percentage of input DNA: % of Input = 2^(CT Input −CT IP )/dilution factor × 100%. j Luciferase reporter assay was performed in isolated WT and KO BMMSCs transfected with pGL3 constructs containing the mouse wild-type Lcn2 promoter (Lcn2-WT) or the mutated Lcn2 promoter with mutations in the NF-κB binding motif (Lcn2-Mut, sequence in supplementary Fig. ). The firefly luciferase activity was measured and normalized to Renilla luciferase activity. k Schematic diagram illustrating the actions of rmCcl2, the CCR2 antagonist, and the NF-κB-specific inhibitor QNZ on NF-κB translocation and Lcn2 expression; created with BioRender ( https://BioRender.com ). l ELISA analysis of total p65 in the nucleus and cytoplasm of BMMSCs. m Representative immunofluorescence images of intracellular p65 localization. Nuclei were stained with DAPI (blue). Scale bar, 50 μm. n P65 nuclear translocation was quantified by ImageJ software via Pearson’s correlation coefficient (PCC) between the p65 immunofluorescence signal and DAPI staining across segmented nuclear regions. o , p WT and KO BMMSCs were treated with the NF-κB-specific inhibitor QNZ (10 nM) for another 2 h after 24-h treatment of rmCcl2 (100 ng/mL) and INCB3344 (10 nM). o The mRNA level of Lcn2 . p Lcn2 levels in the culture medium were determined by ELISA. q Illustration of how the AP-1 inhibitor modulates Ccl2 gene expression; created with BioRender ( https://BioRender.com ). r–u WT and KO BMMSCs were treated with the AP-1-specific inhibitor T-5224 (10 μM) for 24 h. r Sonicated BMMSCs were subjected to ChIP assay using an anti-c-Jun antibody. Representative gel images of end-point ChIP-qPCR products are shown. s Quantitative results are shown as the percentage of input DNA. t The mRNA level of Ccl2 . u Ccl2 levels in the culture medium were determined by ELISA. v Luciferase reporter assay was performed in isolated WT and KO BMMSCs transfected with pGL3 constructs containing the mouse wild-type Ccl2 promoter (Ccl2-WT) or the mutated Ccl2 promoter with mutations in the c-Jun binding motif (Ccl2-Mut, sequence in supplementary Fig. ). The firefly luciferase activity was measured and normalized to Renilla luciferase activity. n = 5 for each group. The data are presented as the means ± SEMs; * p < 0.05, ** p < 0.01, *** p < 0.001. See also supplementary Figs. –

    Article Snippet: The luciferase reporter assay was performed by transfecting 0.2 μg of firefly and Renilla luciferase reporter vectors (Addgene, #128046) driven by either the wild-type mouse Lcn2 or Ccl2 promoter (−1999/+1) or their respective mutants (in which the NF-κB or c-Jun DNA binding site was mutated) (Azenta) into BMMSCs seeded in 96-well plates using Lipofectamine 3000 transfection reagent (Thermo Fisher Scientific, #L3000001).

    Techniques: Expressing, Inhibition, Activation Assay, Isolation, Gene Expression, Real-time Polymerase Chain Reaction, Recombinant, Sonication, Chromatin Immunoprecipitation, Amplification, ChIP-qPCR, Luciferase, Reporter Assay, Transfection, Construct, Binding Assay, Sequencing, Activity Assay, Translocation Assay, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Software