Nanopublications

Matches in Nanopublications for { ?s ?p ?o <http://www.tkuhn.ch/bel2nanopub/RAi3f1gxdIigGw_sFEPRAC_GAzBgXJfsoWmFT1_vzRHS8#provenance>. }

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_6 label "Selventa" provenance.
large_corpus.bel title "BEL Framework Large Corpus Document" provenance.
large_corpus.bel description "Approximately 61,000 statements." provenance.
large_corpus.bel version "20131211" provenance.
large_corpus.bel authoredBy _6 provenance.
assertion wasDerivedFrom large_corpus.bel provenance.
assertion wasDerivedFrom _5 provenance.
assertion hadPrimarySource 16790525 provenance.
_5 wasQuotedFrom 16790525 provenance.
_5 value "we performed cotransfection of Nanog promoter reporter with FoxD3 expression vector in F9 EC cells. As shown in Fig. 2B , FoxD3 activated Nanog promoter in a dose-dependent fashion in F9 cells (Fig. 2B ). To further confirm this finding, we performed the same experiments in ES cells and NIH3T3 cells. As expected, FoxD3 strongly up-regulated the activity of Nanog in ES cells (Fig. 2C ), but surprisingly, had no effect on the same reporter in nonpluripotent NIH3T3 cells (Fig. 2D ). These findings suggest that FoxD3 may activate Nanog in a pluripotency-specific manner. FoxD3 was originally identified as a transcription repressor through binding an AT rich cis-element (20) . To map the cis-elements that FoxD3 utilizes to activate Nanog, a series of deletions were made in Nanog promoter and their activities analyzed. As shown in Fig. 2E , FoxD3 strongly activated reporters bearing –554, –676, and –733 deletions but failed to activate a construct deleting sequences upstream of –269, suggesting that there is a FoxD3 binding site between –269 and –544 in Nanog promoter. Indeed, ChiP analysis confirmed the binding of FoxD3 within this region in vivo in F9 cells (Fig. 2F ). A closer examination of the sequences in the –269 and –554 region identified an AT rich element, which is highly homologous to the identified ES specific enhancer (Fig. 2G, H ). This enhancer was shown as a FoxD3 binding site in vitro (20) . To further investigate this AT element, we inserted three copies of this element upstream of a minimal TK promoter driving a luciferase reporter (Fig. 2H ). When coexpressed with FoxD3, we found that this element is indeed regulated by FoxD3, albeit negatively as reported previously (Fig. 2H ; ref 19 ). Given our finding that FoxD3 activates Nanog expression transcriptionally, we suggest that FoxD3 may assume a dual role in regulating downstream genes, either activation or repression, depending on the context of the target promoter. In the case of Nanog, we showed here that FoxD3 is an activator and, thus, may play an important role in sustaining the expression of Nanog in ES cells." provenance.