Enhancer ID: | E_02_0881 |
Species: | mouse |
Position : | chr1:23881244-23881761 |
Biosample name: | |
Experiment class : | Low throughput |
Enhancer type: | Enhancer |
Disease: | Cardiometabolic disorders |
Pubmed ID: | 25505291 |
Enhancer experiment: | Luciferase Reporter Assay |
Enhancer experiment description: | Using ECR browser (http://ecrbrowser.dcode.org/), we found two conserved regions,ECR1 located between -3177 and -2660 bp upstream of transcription start and ECR2 located between +4517 and 4662 bp downstream of the gene (sequences provided in Supplemental Table 1). ECR1 overlaps with the enhancer described by Shepherd et al. (32), whereas ECR2 has not been described to date. To test the impact of identi?ed ECRs on the regulation of Id3 expression, we cloned them into a pGL3basic vector together with about a 1-kb fragment of the Id3 promoter (Fig. 1A). Obtained plasmids were transfected using nucleofection into primary monocyte-derived macrophages cultivated in the presence of IL-4 and dexamethasone for 6 d. Cells after transfection were further cultivated for 24 h and then stimulated with TGF- 1forthenext 24 h. Cells were harvested and luciferase activity was measured. Without TGF- 1 stimulation, all reporter constructs showed low promoter activity. Stimulation of macrophages with TGF- 1ledto a 2-fold upregulation of the promoter activity. Addition of ECR1 or ECR2 led to a 5-fold increase of promoter activity. Reporter plasmid containing both of them showed nearly 12-fold upregulation of promoter activity, indicating that ECRs have an additive effect (Fig. 1B). |
Target gene : | ID3(HEIR-1,bHLHb25) |
Strong evidence: | -- |
Less strong evidence: | Luciferase Reporter Assay |
Target gene experiment description: | To test the impact of identi?ed ECRs on the regulation of Id3 expression, we cloned them into a pGL3basic vector together with about a 1-kb fragment of the Id3 promoter (Fig. 1A). Obtained plasmids were transfected using nucleofection into primary monocyte-derived macrophages cultivated in the presence of IL-4 and dexamethasone for 6 d. Cells after transfection were further cultivated for 24 h and then stimulated with TGF- 1forthenext 24 h. Cells were harvested and luciferase activity was measured. Without TGF- 1 stimulation, all reporter constructs showed low promoter activity. Stimulation of macrophages with TGF- 1ledto a 2-fold upregulation of the promoter activity. Addition of ECR1 or ECR2 led to a 5-fold increase of promoter activity. Reporter plasmid containing both of them showed nearly 12-fold upregulation of promoter activity, indicating that ECRs have an additive effect (Fig. 1B). |
TF name : | -- |
TF experiment: | -- |
TF experiment description: | -- |
Enhancer function : | -- |
Enhancer function experiment: | -- |
Enhancer function experiment description: |
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SNP ID: | -- |
GeneName | Pathway Name | Source | Gene Number |
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