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pgl2basic vector  (Promega)

 
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    Structured Review

    Promega pgl2basic vector
    ( A ) Sequence of the human hepcidin promoter (acc. no. AD000684.1) with two putative HIF binding motifs which also conform to E-box/USF binding sites (yellow boxes, red letters) and one additional HIF binding site (yellow); binding sites for p53 (light blue), AP1 (blue-green), C/EBPα (green), STAT3 (pink), SMAD (grey) are also marked; mRNA sequence (bold), translation start codon (ATG); underlined sequence represents region highly conserved between human and murine hepcidin 1 gene. Lower sequence, two putative HREs were identified in the mouse hepcidin 1 gene about 2.1 kbp upstream to the transcription start. These HIF binding sites do not conform to E-box/USF binding site consensus sequence. ( B ) A 617-bp human hepcidin promoter construct (HAMP.prom) responded differently to hypoxia (hyp), DMOG and DP in HepG2 and Huh7 cells after 16 h of stimulation (co = control). Data are means±SD of five (HepG2 cells) or three (Huh7 cells) independent experiments. ( C ) SiRNA knock-down of HIF-1α (1α) or HIF-2α (2α) did not reverse the down-regulation of promoter activity by DMOG in Huh7 cells; a 6xHRE luciferase reporter served as control. 3 luc siRNAs served as negative control for the pGL2-based hepcidin promoter constructs and 2 luc siRNA as negative control for the pGL3 6xHRE; deletion of the putative HREs in the HAMP promoter (HAMP.promΔHRE) did not alter the response of the luciferase construct to DMOG nor did HIF-α knock-down. ( D ) Deletion of putative HREs (HAMP.promΔHRE) did not alter the response of the hepcidin promoter to DMOG, DP or hypoxia in Huh7 cells. ( E ) Overexpression of a stable HIF-1α triple mutant (HIF-1αTM) tended to increase hepcidin promoter activity in comparison with the empty vector control (pcDNA3). The 6xHRE reporter served as positive and the promoter-less <t>pGL2basic</t> vector as negative control, respectively; HAMP promoter activities given in B–D are means of three independent experiments±SD. * p <0.05; ** p <0.01 vs. unstimulated control (co).
    Pgl2basic 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
    https://www.bioz.com/result/pgl2basic vector/product/Promega
    Average 90 stars, based on 1 article reviews
    pgl2basic vector - by Bioz Stars, 2026-06
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    Images

    1) Product Images from "Evidence for a Lack of a Direct Transcriptional Suppression of the Iron Regulatory Peptide Hepcidin by Hypoxia-Inducible Factors"

    Article Title: Evidence for a Lack of a Direct Transcriptional Suppression of the Iron Regulatory Peptide Hepcidin by Hypoxia-Inducible Factors

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0007875

    ( A ) Sequence of the human hepcidin promoter (acc. no. AD000684.1) with two putative HIF binding motifs which also conform to E-box/USF binding sites (yellow boxes, red letters) and one additional HIF binding site (yellow); binding sites for p53 (light blue), AP1 (blue-green), C/EBPα (green), STAT3 (pink), SMAD (grey) are also marked; mRNA sequence (bold), translation start codon (ATG); underlined sequence represents region highly conserved between human and murine hepcidin 1 gene. Lower sequence, two putative HREs were identified in the mouse hepcidin 1 gene about 2.1 kbp upstream to the transcription start. These HIF binding sites do not conform to E-box/USF binding site consensus sequence. ( B ) A 617-bp human hepcidin promoter construct (HAMP.prom) responded differently to hypoxia (hyp), DMOG and DP in HepG2 and Huh7 cells after 16 h of stimulation (co = control). Data are means±SD of five (HepG2 cells) or three (Huh7 cells) independent experiments. ( C ) SiRNA knock-down of HIF-1α (1α) or HIF-2α (2α) did not reverse the down-regulation of promoter activity by DMOG in Huh7 cells; a 6xHRE luciferase reporter served as control. 3 luc siRNAs served as negative control for the pGL2-based hepcidin promoter constructs and 2 luc siRNA as negative control for the pGL3 6xHRE; deletion of the putative HREs in the HAMP promoter (HAMP.promΔHRE) did not alter the response of the luciferase construct to DMOG nor did HIF-α knock-down. ( D ) Deletion of putative HREs (HAMP.promΔHRE) did not alter the response of the hepcidin promoter to DMOG, DP or hypoxia in Huh7 cells. ( E ) Overexpression of a stable HIF-1α triple mutant (HIF-1αTM) tended to increase hepcidin promoter activity in comparison with the empty vector control (pcDNA3). The 6xHRE reporter served as positive and the promoter-less pGL2basic vector as negative control, respectively; HAMP promoter activities given in B–D are means of three independent experiments±SD. * p <0.05; ** p <0.01 vs. unstimulated control (co).
    Figure Legend Snippet: ( A ) Sequence of the human hepcidin promoter (acc. no. AD000684.1) with two putative HIF binding motifs which also conform to E-box/USF binding sites (yellow boxes, red letters) and one additional HIF binding site (yellow); binding sites for p53 (light blue), AP1 (blue-green), C/EBPα (green), STAT3 (pink), SMAD (grey) are also marked; mRNA sequence (bold), translation start codon (ATG); underlined sequence represents region highly conserved between human and murine hepcidin 1 gene. Lower sequence, two putative HREs were identified in the mouse hepcidin 1 gene about 2.1 kbp upstream to the transcription start. These HIF binding sites do not conform to E-box/USF binding site consensus sequence. ( B ) A 617-bp human hepcidin promoter construct (HAMP.prom) responded differently to hypoxia (hyp), DMOG and DP in HepG2 and Huh7 cells after 16 h of stimulation (co = control). Data are means±SD of five (HepG2 cells) or three (Huh7 cells) independent experiments. ( C ) SiRNA knock-down of HIF-1α (1α) or HIF-2α (2α) did not reverse the down-regulation of promoter activity by DMOG in Huh7 cells; a 6xHRE luciferase reporter served as control. 3 luc siRNAs served as negative control for the pGL2-based hepcidin promoter constructs and 2 luc siRNA as negative control for the pGL3 6xHRE; deletion of the putative HREs in the HAMP promoter (HAMP.promΔHRE) did not alter the response of the luciferase construct to DMOG nor did HIF-α knock-down. ( D ) Deletion of putative HREs (HAMP.promΔHRE) did not alter the response of the hepcidin promoter to DMOG, DP or hypoxia in Huh7 cells. ( E ) Overexpression of a stable HIF-1α triple mutant (HIF-1αTM) tended to increase hepcidin promoter activity in comparison with the empty vector control (pcDNA3). The 6xHRE reporter served as positive and the promoter-less pGL2basic vector as negative control, respectively; HAMP promoter activities given in B–D are means of three independent experiments±SD. * p <0.05; ** p <0.01 vs. unstimulated control (co).

    Techniques Used: Sequencing, Binding Assay, Construct, Activity Assay, Luciferase, Negative Control, Over Expression, Mutagenesis, Plasmid Preparation



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    ( A ) Sequence of the human hepcidin promoter (acc. no. AD000684.1) with two putative HIF binding motifs which also conform to E-box/USF binding sites (yellow boxes, red letters) and one additional HIF binding site (yellow); binding sites for p53 (light blue), AP1 (blue-green), C/EBPα (green), STAT3 (pink), SMAD (grey) are also marked; mRNA sequence (bold), translation start codon (ATG); underlined sequence represents region highly conserved between human and murine hepcidin 1 gene. Lower sequence, two putative HREs were identified in the mouse hepcidin 1 gene about 2.1 kbp upstream to the transcription start. These HIF binding sites do not conform to E-box/USF binding site consensus sequence. ( B ) A 617-bp human hepcidin promoter construct (HAMP.prom) responded differently to hypoxia (hyp), DMOG and DP in HepG2 and Huh7 cells after 16 h of stimulation (co = control). Data are means±SD of five (HepG2 cells) or three (Huh7 cells) independent experiments. ( C ) SiRNA knock-down of HIF-1α (1α) or HIF-2α (2α) did not reverse the down-regulation of promoter activity by DMOG in Huh7 cells; a 6xHRE luciferase reporter served as control. 3 luc siRNAs served as negative control for the pGL2-based hepcidin promoter constructs and 2 luc siRNA as negative control for the pGL3 6xHRE; deletion of the putative HREs in the HAMP promoter (HAMP.promΔHRE) did not alter the response of the luciferase construct to DMOG nor did HIF-α knock-down. ( D ) Deletion of putative HREs (HAMP.promΔHRE) did not alter the response of the hepcidin promoter to DMOG, DP or hypoxia in Huh7 cells. ( E ) Overexpression of a stable HIF-1α triple mutant (HIF-1αTM) tended to increase hepcidin promoter activity in comparison with the empty vector control (pcDNA3). The 6xHRE reporter served as positive and the promoter-less <t>pGL2basic</t> vector as negative control, respectively; HAMP promoter activities given in B–D are means of three independent experiments±SD. * p <0.05; ** p <0.01 vs. unstimulated control (co).
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    ( A ) Sequence of the human hepcidin promoter (acc. no. AD000684.1) with two putative HIF binding motifs which also conform to E-box/USF binding sites (yellow boxes, red letters) and one additional HIF binding site (yellow); binding sites for p53 (light blue), AP1 (blue-green), C/EBPα (green), STAT3 (pink), SMAD (grey) are also marked; mRNA sequence (bold), translation start codon (ATG); underlined sequence represents region highly conserved between human and murine hepcidin 1 gene. Lower sequence, two putative HREs were identified in the mouse hepcidin 1 gene about 2.1 kbp upstream to the transcription start. These HIF binding sites do not conform to E-box/USF binding site consensus sequence. ( B ) A 617-bp human hepcidin promoter construct (HAMP.prom) responded differently to hypoxia (hyp), DMOG and DP in HepG2 and Huh7 cells after 16 h of stimulation (co = control). Data are means±SD of five (HepG2 cells) or three (Huh7 cells) independent experiments. ( C ) SiRNA knock-down of HIF-1α (1α) or HIF-2α (2α) did not reverse the down-regulation of promoter activity by DMOG in Huh7 cells; a 6xHRE luciferase reporter served as control. 3 luc siRNAs served as negative control for the pGL2-based hepcidin promoter constructs and 2 luc siRNA as negative control for the pGL3 6xHRE; deletion of the putative HREs in the HAMP promoter (HAMP.promΔHRE) did not alter the response of the luciferase construct to DMOG nor did HIF-α knock-down. ( D ) Deletion of putative HREs (HAMP.promΔHRE) did not alter the response of the hepcidin promoter to DMOG, DP or hypoxia in Huh7 cells. ( E ) Overexpression of a stable HIF-1α triple mutant (HIF-1αTM) tended to increase hepcidin promoter activity in comparison with the empty vector control (pcDNA3). The 6xHRE reporter served as positive and the promoter-less <t>pGL2basic</t> vector as negative control, respectively; HAMP promoter activities given in B–D are means of three independent experiments±SD. * p <0.05; ** p <0.01 vs. unstimulated control (co).
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    Image Search Results


    ( A ) Sequence of the human hepcidin promoter (acc. no. AD000684.1) with two putative HIF binding motifs which also conform to E-box/USF binding sites (yellow boxes, red letters) and one additional HIF binding site (yellow); binding sites for p53 (light blue), AP1 (blue-green), C/EBPα (green), STAT3 (pink), SMAD (grey) are also marked; mRNA sequence (bold), translation start codon (ATG); underlined sequence represents region highly conserved between human and murine hepcidin 1 gene. Lower sequence, two putative HREs were identified in the mouse hepcidin 1 gene about 2.1 kbp upstream to the transcription start. These HIF binding sites do not conform to E-box/USF binding site consensus sequence. ( B ) A 617-bp human hepcidin promoter construct (HAMP.prom) responded differently to hypoxia (hyp), DMOG and DP in HepG2 and Huh7 cells after 16 h of stimulation (co = control). Data are means±SD of five (HepG2 cells) or three (Huh7 cells) independent experiments. ( C ) SiRNA knock-down of HIF-1α (1α) or HIF-2α (2α) did not reverse the down-regulation of promoter activity by DMOG in Huh7 cells; a 6xHRE luciferase reporter served as control. 3 luc siRNAs served as negative control for the pGL2-based hepcidin promoter constructs and 2 luc siRNA as negative control for the pGL3 6xHRE; deletion of the putative HREs in the HAMP promoter (HAMP.promΔHRE) did not alter the response of the luciferase construct to DMOG nor did HIF-α knock-down. ( D ) Deletion of putative HREs (HAMP.promΔHRE) did not alter the response of the hepcidin promoter to DMOG, DP or hypoxia in Huh7 cells. ( E ) Overexpression of a stable HIF-1α triple mutant (HIF-1αTM) tended to increase hepcidin promoter activity in comparison with the empty vector control (pcDNA3). The 6xHRE reporter served as positive and the promoter-less pGL2basic vector as negative control, respectively; HAMP promoter activities given in B–D are means of three independent experiments±SD. * p <0.05; ** p <0.01 vs. unstimulated control (co).

    Journal: PLoS ONE

    Article Title: Evidence for a Lack of a Direct Transcriptional Suppression of the Iron Regulatory Peptide Hepcidin by Hypoxia-Inducible Factors

    doi: 10.1371/journal.pone.0007875

    Figure Lengend Snippet: ( A ) Sequence of the human hepcidin promoter (acc. no. AD000684.1) with two putative HIF binding motifs which also conform to E-box/USF binding sites (yellow boxes, red letters) and one additional HIF binding site (yellow); binding sites for p53 (light blue), AP1 (blue-green), C/EBPα (green), STAT3 (pink), SMAD (grey) are also marked; mRNA sequence (bold), translation start codon (ATG); underlined sequence represents region highly conserved between human and murine hepcidin 1 gene. Lower sequence, two putative HREs were identified in the mouse hepcidin 1 gene about 2.1 kbp upstream to the transcription start. These HIF binding sites do not conform to E-box/USF binding site consensus sequence. ( B ) A 617-bp human hepcidin promoter construct (HAMP.prom) responded differently to hypoxia (hyp), DMOG and DP in HepG2 and Huh7 cells after 16 h of stimulation (co = control). Data are means±SD of five (HepG2 cells) or three (Huh7 cells) independent experiments. ( C ) SiRNA knock-down of HIF-1α (1α) or HIF-2α (2α) did not reverse the down-regulation of promoter activity by DMOG in Huh7 cells; a 6xHRE luciferase reporter served as control. 3 luc siRNAs served as negative control for the pGL2-based hepcidin promoter constructs and 2 luc siRNA as negative control for the pGL3 6xHRE; deletion of the putative HREs in the HAMP promoter (HAMP.promΔHRE) did not alter the response of the luciferase construct to DMOG nor did HIF-α knock-down. ( D ) Deletion of putative HREs (HAMP.promΔHRE) did not alter the response of the hepcidin promoter to DMOG, DP or hypoxia in Huh7 cells. ( E ) Overexpression of a stable HIF-1α triple mutant (HIF-1αTM) tended to increase hepcidin promoter activity in comparison with the empty vector control (pcDNA3). The 6xHRE reporter served as positive and the promoter-less pGL2basic vector as negative control, respectively; HAMP promoter activities given in B–D are means of three independent experiments±SD. * p <0.05; ** p <0.01 vs. unstimulated control (co).

    Article Snippet: Three different hepcidin promoter fragments were amplified from genomic DNA by PCR using the primers given in and Combizyme Polymerase Mix (Invitek), and cloned into the pGL2basic vector (Promega).

    Techniques: Sequencing, Binding Assay, Construct, Activity Assay, Luciferase, Negative Control, Over Expression, Mutagenesis, Plasmid Preparation