PMC:2783367 / 37155-38700
Annnotations
{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/2783367","sourcedb":"PMC","sourceid":"2783367","source_url":"https://www.ncbi.nlm.nih.gov/pmc/2783367","text":"Nuclear β-catenin forms a complex with the transcription factor TCF/LEF to bind to the promoter of target genes and to stimulate target gene transcription 42, 43. EMSA were conducted to further verify the effect of curcumin on interrupting canonical Wnt signaling. Nuclear protein extracts from HSCs treated with curcumin at 0-30 μM for 24 hr were incubated with biotin-labeled double-stranded oligonucleotide probe P(TCFwt). The probe P(TCFwt) contained the consensus TCF/LEF binding site found in the lox-1 promoter. As shown in Fig. 9C, curcumin dose-dependently reduced the DNA binding activity of the transcription factor TCF/LEF to the probe. To examine the DNA binding specificity of TCF/LEF to the probe, competition assays were performed using a 10-, 50-, or 100-fold excess of the unlabeled probe P(TCFwt) (lanes 3-5 in Fig. 9D), or the unlabeled probe P(TCFmut) (lanes 6-8 in Fig. 9D). The probe P(TCFmut) contained the TCF/LEF binding site found in the lox-1 promoter with site-directed mutations. It was found that the amount of TCF/LEF binding to the biotin-labeled probe P(TCFwt)) was competitively reduced by the unlabeled probe P(TCFwt), but not by the mutant probe P(TCFwt). These results indicated that curcumin dose-dependently reduced the DNA binding activity of TCF/LEF to the promoter of LOX-1 gene in activated HSCs. Taken together, these results indicated that curcumin interrupted canonical Wnt signaling in activated HSCs by reducing the abundance of nuclear β-catenin and reducing the DNA binding activity of TCF/LEF.","tracks":[{"project":"2_test","denotations":[{"id":"19736547-18177486-66178374","span":{"begin":155,"end":157},"obj":"18177486"},{"id":"19736547-18177486-66178374","span":{"begin":155,"end":157},"obj":"18177486"},{"id":"19736547-18160255-66178375","span":{"begin":159,"end":161},"obj":"18160255"},{"id":"19736547-18160255-66178375","span":{"begin":159,"end":161},"obj":"18160255"},{"id":"T51487","span":{"begin":155,"end":157},"obj":"18177486"},{"id":"T55603","span":{"begin":155,"end":157},"obj":"18177486"},{"id":"T45850","span":{"begin":159,"end":161},"obj":"18160255"},{"id":"T56336","span":{"begin":159,"end":161},"obj":"18160255"}],"attributes":[{"subj":"19736547-18177486-66178374","pred":"source","obj":"2_test"},{"subj":"19736547-18177486-66178374","pred":"source","obj":"2_test"},{"subj":"19736547-18160255-66178375","pred":"source","obj":"2_test"},{"subj":"19736547-18160255-66178375","pred":"source","obj":"2_test"},{"subj":"T51487","pred":"source","obj":"2_test"},{"subj":"T55603","pred":"source","obj":"2_test"},{"subj":"T45850","pred":"source","obj":"2_test"},{"subj":"T56336","pred":"source","obj":"2_test"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"2_test","color":"#93ece2","default":true}]}]}}