دورية أكاديمية
Image-based high-throughput mapping of TGF-β-induced phosphocomplexes at a single-cell level.
| العنوان: | Image-based high-throughput mapping of TGF-β-induced phosphocomplexes at a single-cell level. |
|---|---|
| المؤلفون: | Lönn P; Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden. peter.loenn@gmail.com., Al-Amin RA; Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden., Doulabi EM; Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden., Heldin J; Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden.; Department of Pharmaceutical Biosciences, Uppsala University, Uppsala, Sweden., Gallini R; Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden., Björkesten J; Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden., Oelrich J; Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden., Kamali-Moghaddam M; Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden., Landegren U; Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden. ulf.landegren@igp.uu.se. |
| المصدر: | Communications biology [Commun Biol] 2021 Nov 12; Vol. 4 (1), pp. 1284. Date of Electronic Publication: 2021 Nov 12. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Nature Publishing Group UK Country of Publication: England NLM ID: 101719179 Publication Model: Electronic Cited Medium: Internet ISSN: 2399-3642 (Electronic) Linking ISSN: 23993642 NLM ISO Abbreviation: Commun Biol Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: London, United Kingdom : Nature Publishing Group UK, [2018]- |
| مواضيع طبية MeSH: | Image Processing, Computer-Assisted* , Protein Interaction Mapping*, Smad4 Protein/*genetics , Transforming Growth Factor beta1/*genetics, HaCaT Cells ; Humans ; Phosphorylation ; Single-Cell Analysis ; Smad4 Protein/metabolism ; Transforming Growth Factor beta1/metabolism |
| مستخلص: | Protein interactions and posttranslational modifications orchestrate cellular responses to e.g. cytokines and drugs, but it has been difficult to monitor these dynamic events in high-throughput. Here, we describe a semi-automated system for large-scale in situ proximity ligation assays (isPLA), combining isPLA in microtiter wells with automated microscopy and computer-based image analysis. Phosphorylations and interactions are digitally recorded along with subcellular morphological features. We investigated TGF-β-responsive Smad2 linker phosphorylations and complex formations over time and across millions of individual cells, and we relate these events to cell cycle progression and local cell crowding via measurements of DNA content and nuclear size of individual cells, and of their relative positions. We illustrate the suitability of this protocol to screen for drug effects using phosphatase inhibitors. Our approach expands the scope for image-based single cell analyses by combining observations of protein interactions and modifications with morphological details of individual cells at high throughput. (© 2021. The Author(s).) |
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| معلومات مُعتمدة: | 2012-5852 Vetenskapsrådet (Swedish Research Council); 2017-04152 Vetenskapsrådet (Swedish Research Council); 2018-02943 Vetenskapsrådet (Swedish Research Council); 2018-06156 Vetenskapsrådet (Swedish Research Council); SB16-0046 Stiftelsen för Strategisk Forskning (Swedish Foundation for Strategic Research); M130/16 Torsten Söderbergs Stiftelse (Torsten Söderberg Foundation); 19 0384 Pj Cancerfonden (Swedish Cancer Society); FP/2007-2013)/ERC Grant Agreement no. 294409 EC | EC Seventh Framework Programm | FP7 Ideas: European Research Council (FP7-IDEAS-ERC - Specific Programme: "Ideas" Implementing the Seventh Framework Programme of the European Community for Research, Technological Development and Demonstration Activities (2007 to 2013)) |
| المشرفين على المادة: | 0 (SMAD4 protein, human) 0 (Smad4 Protein) 0 (TGFB1 protein, human) 0 (Transforming Growth Factor beta1) |
| تواريخ الأحداث: | Date Created: 20211113 Date Completed: 20211220 Latest Revision: 20211220 |
| رمز التحديث: | 20231215 |
| مُعرف محوري في PubMed: | PMC8590043 |
| DOI: | 10.1038/s42003-021-02798-4 |
| PMID: | 34773084 |
| قاعدة البيانات: | MEDLINE |
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