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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/RA_M4pTNu7Su-05_AVyqGeQHvT9lwFwjiUYy9WWUnbhso#_6 http://resource.belframework.org/belframework/20131211/knowledge/large_corpus.bel http://purl.org/pav/version 20131211 http://www.tkuhn.ch/bel2nanopub/RA_M4pTNu7Su-05_AVyqGeQHvT9lwFwjiUYy9WWUnbhso#_5 http://www.w3.org/ns/prov#value ErbB4 downregulation in fetal rat type II epithelial cells. Primary fetal rat type II cells were transfected with three different siRNAs targeting three different sequences in the transcribed erbB4 mRNA, a methodology that has been used by others (12). To identify the most effective dose of siRNA in these primary fetal type II cells, we performed a dose-response curve and found that concentrations of 60 nM and 120 nM showed similar effects with a dose-dependent U-shaped response curve (Fig. 1A). This U-shaped response has been documented for some genes, is not related to the sequence of siRNA used, and starts at ~100 ng of siRNA (Dr. Susan Magdaleno, Ambion, Austin, TX, personal communication). To minimize toxicity while optimizing the siRNA inhibitory effect, we chose the dose for subsequent experiments that was midrange between these doses at 90 nM (30 nM of each sequence). The transfection efficiency using this concentration, observed by fluorescein labeling of the siRNA, was 71%. Densitometry (Fig. 1B) of eight experiments using the combination of three different sequences against erbB4 at a concentration of 30 nM each showed a significant decrease in erbB4 protein to 74 ± 2.5% (means ± SE, n = 8, P < 0.0001) compared with scrambled siRNA.Endogenous baseline tyrosine phosphorylation (as an indicator of ERBB4 activity) was reduced to 81 ± 8% (means ± SE, n = 4, P = 0.05) with the same concentration of erbB4 siRNA treatment compared with scrambled siRNA treatment. NRG-induced phosphorylation was reduced to 80 ± 15% (means ± SE, n = 4, P = 0.17), and FCM-induced phosphorylation was significantly reduced to 73 ± 12% (means ± SE, n = 4, P = 0.029) by erbB4 siRNA treatment (Fig. 1C). http://www.tkuhn.ch/bel2nanopub/RA_M4pTNu7Su-05_AVyqGeQHvT9lwFwjiUYy9WWUnbhso#_5 http://www.w3.org/ns/prov#wasQuotedFrom http://www.ncbi.nlm.nih.gov/pubmed/17545485 http://www.tkuhn.ch/bel2nanopub/RA_M4pTNu7Su-05_AVyqGeQHvT9lwFwjiUYy9WWUnbhso#_6 http://www.w3.org/2000/01/rdf-schema#label Selventa http://www.tkuhn.ch/bel2nanopub/RA_M4pTNu7Su-05_AVyqGeQHvT9lwFwjiUYy9WWUnbhso#assertion http://www.w3.org/ns/prov#hadPrimarySource http://www.ncbi.nlm.nih.gov/pubmed/17545485 http://www.tkuhn.ch/bel2nanopub/RA_M4pTNu7Su-05_AVyqGeQHvT9lwFwjiUYy9WWUnbhso#assertion http://www.w3.org/ns/prov#wasDerivedFrom http://resource.belframework.org/belframework/20131211/knowledge/large_corpus.bel http://www.tkuhn.ch/bel2nanopub/RA_M4pTNu7Su-05_AVyqGeQHvT9lwFwjiUYy9WWUnbhso#assertion http://www.w3.org/ns/prov#wasDerivedFrom http://www.tkuhn.ch/bel2nanopub/RA_M4pTNu7Su-05_AVyqGeQHvT9lwFwjiUYy9WWUnbhso#_5 http://www.tkuhn.ch/bel2nanopub/RA_M4pTNu7Su-05_AVyqGeQHvT9lwFwjiUYy9WWUnbhso#pubinfo http://www.tkuhn.ch/bel2nanopub/RA_M4pTNu7Su-05_AVyqGeQHvT9lwFwjiUYy9WWUnbhso http://purl.org/dc/terms/created 2014-07-03T14:33:10.845+02:00 http://www.tkuhn.ch/bel2nanopub/RA_M4pTNu7Su-05_AVyqGeQHvT9lwFwjiUYy9WWUnbhso http://purl.org/pav/createdBy http://orcid.org/0000-0001-6818-334X http://www.tkuhn.ch/bel2nanopub/RA_M4pTNu7Su-05_AVyqGeQHvT9lwFwjiUYy9WWUnbhso http://purl.org/pav/createdBy http://orcid.org/0000-0002-1267-0234