About Enhancer
| Enhancer ID: | E_02_0792 |
| Species: | human |
| Position : | chr3:181432912-181433312   |
| Biosample name: | |
| Experiment class : | low throughput |
| Enhancer type: | Enhancer |
| Disease: | Lung cancer |
| Pubmed ID: | 28737489 |
| Enhancer experiment: | Luciferase Reporter Assay,ChIP-qPCR |
| Enhancer experiment description: | To further delineate the molecular mechanism of SOX2 regulation by NFATc2, we screened, in silico, the genomic sequences spanning 5 kb up- and downstream of the SOX2 transcription start site (TSS), which identified 4 regions encompassing multiple conserved NFAT binding sequences (Figure 6-figure supplement 1A). Luciferase assays confirmed these regions are active transcriptional regulatory regions (Figure 6-figure supplement 1B). Further evaluation with respective SOX2 luciferase reporter revealed transcriptional activities were mediated by sites 1, 2, 4, and 5 (Figure 6B). Using H441 lung cancer cell line with NFATc2 transient overexpression, we observed only sites 1, 4 and 5 showed statistically significant increased reporter activities while those of sites 4 and 5 were reciprocally abolished by CSA treatment (Figure 6C). |
About Target gene
| Target gene : | SOX2 (ANOP3,MCOPS3) |
| Strong evidence: | -- |
| Less strong evidence: | Luciferase Reporter Assay,ChIP-qPCR |
| Target gene experiment description: | To further delineate the molecular mechanism of SOX2 regulation by NFATc2, we screened, in silico, the genomic sequences spanning 5 kb up- and downstream of the SOX2 transcription start site (TSS), which identified 4 regions encompassing multiple conserved NFAT binding sequences (Figure 6-figure supplement 1A). Luciferase assays confirmed these regions are active transcriptional regulatory regions (Figure 6-figure supplement 1B). Further evaluation with respective SOX2 luciferase reporter revealed transcriptional activities were mediated by sites 1, 2, 4, and 5 (Figure 6B). Using H441 lung cancer cell line with NFATc2 transient overexpression, we observed only sites 1, 4 and 5 showed statistically significant increased reporter activities while those of sites 4 and 5 were reciprocally abolished by CSA treatment (Figure 6C). |
About TF
| TF name : | NFATC2(NFAT1,NFATP) |
| TF experiment: | Luciferase Reporter Assay,ChIP-qPCR |
| TF experiment description: | Finally, site directed mutagenesis of NFAT motifs (GGAAA to GACTA) prevented reporter activities of sites 4 and 5 only (Figure 6D), and the findings were supported by data from A549 and H1299 cells ectopically expressing NFATc2, respectively (Figure 6-figure supplement 1C,D). Thus, the data suggested NFATc2 was highly likely to regulate SOX2 expression through binding to 3 enhancers at sites 4 and 5. For validation, NFATc2 ChIP-qPCR assays were performed using A549 with NFATc2 upregulation, which showed statistically significant enrichment of sites 4 and 5 sequences compared to vector control (Figure 6F). In HCC827 cells, sites 4 and 5 sequences were significantly enriched by anti-NFATc2 antibody compared to IgG control. Conversely, these sequences were significantly reduced upon NFATc2 knockout in HCC827, compared to their endogenous levels in control cells, indicating de novo physical binding of NFATc2 to SOX2 at sites 4 and 5 (Figure 6G). Together, the data showed NFATc2 upregulates SOX2 by binding to its 3 enhancer region at around 3.2 kb (site 4) and 3.6 kb (site 5) from the TSS, respectively |
About Function
| Enhancer function : | -- |
| Enhancer function experiment: | -- |
| Enhancer function experiment description: |
-- |
About SNP
| SNP ID: | -- |
Upstream Pathway Annotation of TF
Enhancer associated network
The number on yellow line represents the distance between enhancer and
target gene