title: Comprehensive analysis of signal transduction in three-dimensional ECM-based tumor cell cultures.
abstract: Analysis of signal transduction and protein phosphorylation is fundamental to understanding physiological and pathological cell behavior and identifying novel therapeutic targets. Despite the fact that the use of physiological three-dimensional cell culture assays is increasing, 3D proteomics and phosphoproteomics remain challenging due to difficulties with easy, robust and reproducible sample preparation. Here, we present an easy-to-perform, reliable and time-efficient method for the production of 3D cell lysates that does not compromise cell adhesion before cell lysis. The samples can be used for western blotting as well as phosphoproteome array technology. This technique will be of interest for researchers working in all fields of biology and drug development.
authors.affiliation: OncoRay - National Center for Radiation Research in Oncology, Medical Faculty Carl Gustav Carus, Dresden University of Technology, 01307 Dresden, Germany ; Radiation Oncology Branch, Center for Cancer Research, National Institutes of Health/National Cancer Institute, Bethesda, MD 20892, USA.
authors.forename: Iris
authors.initials: I
authors.lastname: Eke
authors.affiliation: OncoRay - National Center for Radiation Research in Oncology, Medical Faculty Carl Gustav Carus, Dresden University of Technology, 01307 Dresden, Germany ; Department of Radiotherapy and Oncology, University of Frankfurt, 60590 Frankfurt am Main, Germany.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Hehlgans
authors.affiliation: Full Moon BioSystems Inc., Sunnyvale, CA 94085, USA.
authors.forename: Yaping
authors.initials: Y
authors.lastname: Zong
authors.affiliation: OncoRay - National Center for Radiation Research in Oncology, Medical Faculty Carl Gustav Carus, Dresden University of Technology, 01307 Dresden, Germany ; Department of Radiation Oncology, University Hospital and Medical Faculty Carl Gustav Carus, Dresden University of Technology, 01307 Dresden, Germany ; Helmholtz Center Dresden-Rossendorf, Institute of Radiooncology, 01328 Dresden, Germany ; German Cancer Consortium (DKTK), 01307 Dresden, Germany ; German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany.
authors.forename: Nils
authors.initials: N
authors.lastname: Cordes
keywords: 3D cell culture; extracellular matrix; phosphoproteomics; protein analysis; signaling