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General Information info

Manuscript title Network quantification of EGFR signaling unveils potential for targeted combination therapy.
PubMed ID 23752269
Journal Molecular Systems Biology
Year 2013
Authors Bertram Klinger, Anja Sieber, Raphaela Fritsche-Guenther, Franziska Witzel, Leanne Berry, Dirk Schumacher, Yibing Yan, Pawel Durek1, Mark Merchant, Reinhold Schaefer, Christine Sers and Nils Bluethgen
Affiliations Laboratory of Molecular Tumour Pathology, Institute of Pathology, Charité - Universitatsmedizin Berlin, Berlin, Germany, and Institute for Theoretical Biology, Humboldt University Berlin, Berlin, Germany
Keywords cancer, EGFR signaling, modular response analysis, signal transduction
Full text article Downloadarticle Klinger_2013.pdf
Project name not specified

Experiment Description info

Organism Homo sapiens
Strain colorectal cancer cell lines SW480, SW403, HCT116, RKO, LIM1215 and HT29
Data type enzyme/protein concentrations
Data units (a.u.)
Execution date not specified

Experimental Details info

Temperature (0C) 37.0
pH not specified
Carbon source not specified
Culture mode batch
Process condition aerobic
Dilution rate (h-1)
Working volume (L) not specified
Biomass concentration (g/L) not specified
Medium composition

All cell lines were maintained in DMEM (Dulbecco’s modified Eagle’s medium, Lonza) supplemented with 10% fetal calf serum, 1% ultraglutamine and 1% penicillin/streptomycin and incubated in a humidified atmosphere of 5% CO2 in air at 37 ºC.

General protocol information Measurement method: immunoblotting

Methods description - Notes

Immunoblotting - protein extracts of cells were prepared as described for Bioplex analysis. Blotting procedure and materials were as previously described [1,2]. The following primary antibodies were used: rabbit anti-human P-p70S6 (Thr389, Cell Signaling Technology, 1:500) or mouse antihuman GAPDH (Ambion, 1:12 500). Membranes were scanned using Li-COR Odyssey. The signals were quantified using Odyssey software.

-------------------References-------------------
[1] Fritsche-Guenther R, Witzel F, Sieber A, Herr R, Schmidt N, Braun S, Brummer T, Sers C, Bluthgen N (2011). Strong negative feedback from Erk to Raf confers robustness to MAPK signalling. Mol Syst Biol 7: 489. http://doi.org/dvbp78
[2] Stelniec-Klotz I, Legewie S, Tchernitsa O, Witzel F, Klinger B, Sers C, Herzel H, Bluthgen N, Schafer R (2012). Reverse engineering a hierarchical regulatory network downstream of oncogenic KRAS. Mol Syst Biol 8: 601. http://doi.org/f2phtg

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Submission and curation info

Entered by Administrator KiMoSysFirst name: Administrator
Affiliation: INESC-ID/IST
Homepage: http://kdbio.inesc-id.pt/kimosys
Interests: mathematical modeling, accessible data, use of data

Created 2014-04-23 16:07:23 UTC

Updated 2014-06-13 23:39:15 UTC

Version 0

Status (reviewed) 2018-07-07 21:37:41 UTC




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