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All rights reserved. http://resource.belframework.org/belframework/20131211/knowledge/large_corpus.bel http://purl.org/dc/elements/1.1/title BEL Framework Large Corpus Document http://resource.belframework.org/belframework/20131211/knowledge/large_corpus.bel http://purl.org/pav/authoredBy http://www.tkuhn.ch/bel2nanopub/RAsjTTzEIaE1x30dKzPwzVpE1-gIq38ThIlDvHTtobkCQ#_6 http://resource.belframework.org/belframework/20131211/knowledge/large_corpus.bel http://purl.org/pav/version 20131211 http://www.tkuhn.ch/bel2nanopub/RAsjTTzEIaE1x30dKzPwzVpE1-gIq38ThIlDvHTtobkCQ#_5 http://www.w3.org/ns/prov#value To investigate whether IGF-1 also enhanced transactivation potential, a reporter gene assay was performed in HEK293 cells. We used a reporter construct incorporating multimeric Lef/Tcf promoter sequences upstream of a luciferase reporter (TOPFLASH) and a control construct containing mutated promoter sequences (FOPFLASH, ref. 21). We cotransfected TOPFLASH or FOPFLASH, ?-catenin, and human wild-type Tcf-4 wild-type expression constructs into 293 cells. Optimized luciferase assays showed a basal level of TOP/FOP activity of between 4:1 and 20:1. Forty-eight hours after transfection, serum-starved 293 cells were treated with IGF-1 or insulin for 3 or 6 h. These factors alone failed to alter basal reporter activity (Fig. 6 b). A 16-h pretreatment of the transfectants with LiCl had a dramatic effect on reporter activity, with significant enhancement at 50 mM LiCl (P < 0.01 by Tukey's test for comparison with activity in the absence of LiCl). We saw a further increase in reporter activity when we added 50 ng/ml (6.5 nM) IGF-1 to transfectants pretreated with LiCl at 10 mM (P < 0.01 after 6 h of IGF-1 treatment) and at 50 mM (P < 0.01 after 3 h of IGF-1 treatment and P < 0.001 after 6 h). http://www.tkuhn.ch/bel2nanopub/RAsjTTzEIaE1x30dKzPwzVpE1-gIq38ThIlDvHTtobkCQ#_5 http://www.w3.org/ns/prov#wasQuotedFrom http://www.ncbi.nlm.nih.gov/pubmed/11035789 http://www.tkuhn.ch/bel2nanopub/RAsjTTzEIaE1x30dKzPwzVpE1-gIq38ThIlDvHTtobkCQ#_6 http://www.w3.org/2000/01/rdf-schema#label Selventa http://www.tkuhn.ch/bel2nanopub/RAsjTTzEIaE1x30dKzPwzVpE1-gIq38ThIlDvHTtobkCQ#assertion http://www.w3.org/ns/prov#hadPrimarySource http://www.ncbi.nlm.nih.gov/pubmed/11035789 http://www.tkuhn.ch/bel2nanopub/RAsjTTzEIaE1x30dKzPwzVpE1-gIq38ThIlDvHTtobkCQ#assertion http://www.w3.org/ns/prov#wasDerivedFrom http://resource.belframework.org/belframework/20131211/knowledge/large_corpus.bel http://www.tkuhn.ch/bel2nanopub/RAsjTTzEIaE1x30dKzPwzVpE1-gIq38ThIlDvHTtobkCQ#assertion http://www.w3.org/ns/prov#wasDerivedFrom http://www.tkuhn.ch/bel2nanopub/RAsjTTzEIaE1x30dKzPwzVpE1-gIq38ThIlDvHTtobkCQ#_5 http://www.tkuhn.ch/bel2nanopub/RAsjTTzEIaE1x30dKzPwzVpE1-gIq38ThIlDvHTtobkCQ#pubinfo http://www.tkuhn.ch/bel2nanopub/RAsjTTzEIaE1x30dKzPwzVpE1-gIq38ThIlDvHTtobkCQ http://purl.org/dc/terms/created 2014-07-03T14:31:37.632+02:00 http://www.tkuhn.ch/bel2nanopub/RAsjTTzEIaE1x30dKzPwzVpE1-gIq38ThIlDvHTtobkCQ http://purl.org/pav/createdBy http://orcid.org/0000-0001-6818-334X http://www.tkuhn.ch/bel2nanopub/RAsjTTzEIaE1x30dKzPwzVpE1-gIq38ThIlDvHTtobkCQ http://purl.org/pav/createdBy http://orcid.org/0000-0002-1267-0234