Search: onr:"swepub:oai:DiVA.org:uu-374248" >
High-throughput in ...
High-throughput in situ mapping of phosphorylated protein complexes across the cell cycle and in response to drugs
-
- Lönn, Peter (author)
- Uppsala universitet,Institutionen för immunologi, genetik och patologi,Science for Life Laboratory, SciLifeLab, Science for Life Laboratory, SciLifeLab,Molekylära verktyg, Molecular tools
-
- Al-Amin, Rasel A., 1983- (author)
- Uppsala universitet,Institutionen för immunologi, genetik och patologi,Science for Life Laboratory, SciLifeLab, Science for Life Laboratory, SciLifeLab,Molekylära verktyg, Molecular tools
-
- Heldin, Johan (author)
- Uppsala universitet,Science for Life Laboratory, SciLifeLab,Institutionen för immunologi, genetik och patologi,Institutionen för farmaceutisk biovetenskap,Molekylära verktyg, Molecular tools
-
show more...
-
- Gallini, Radiosa (author)
- Uppsala universitet,Institutionen för immunologi, genetik och patologi,Science for Life Laboratory, SciLifeLab, Science for Life Laboratory, SciLifeLab,Molekylära verktyg, Molecular tools
-
- Björkesten, Johan (author)
- Uppsala universitet,Science for Life Laboratory, SciLifeLab,Institutionen för immunologi, genetik och patologi,Molekylära verktyg, Molecular tools
-
- Oelrich, Johan (author)
- Uppsala universitet,Molekylära verktyg,Science for Life Laboratory, SciLifeLab, Science for Life Laboratory, SciLifeLab,Molekylära verktyg, Molecular tools
-
- Kamali-Moghaddam, Masood (author)
- Uppsala universitet,Molekylära verktyg,Science for Life Laboratory, SciLifeLab,Institutionen för medicinska vetenskaper
-
- Landegren, Ulf (author)
- Uppsala universitet,Molekylära verktyg,Science for Life Laboratory, SciLifeLab
-
show less...
-
(creator_code:org_t)
- English.
- Related links:
-
https://urn.kb.se/re...
Abstract
Subject headings
Close
- Interactions and posttranslational modifications (PTMs) of proteins orchestrate cellular responses to cytokines, drugs or other agents, but it has been difficult to monitor and characterize these dynamic events at high-throughput. Here, we have established a semi-automated system for large-scale in situ proximity ligation assays (isPLA). The protocol combines isPLA in microtiter wells with automated microscopy and computer-based image analysis whereby specific protein phosphorylations and interactions are digitally recorded in cells, along with measurements of morphological features. We demonstrate how this platform can improve analysis of cellular signaling by investigating TGF-b responsive Smad2 linker phosphorylations and complex formations over time and across millions of individual cells. We depict single cell responses in relation to e.g. local cell crowding and cell cycle progression via measurements of DNA content and nuclear size. Finally, we illustrate the application of the protocol for demonstrating drug effects by screening a library of phosphatase inhibitors. In summary, 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.
Keyword
- Biokemi
- Biochemistry
- Molekylär cellbiologi
- Molecular Cellbiology
- Molecular Biotechnology
- Molekylär bioteknik
Publication and Content Type
- vet (subject category)
- ovr (subject category)
To the university's database