ArticleAnalytical biochemistry2019
Quantitative in situ proximity ligation assays examining protein interactions and phosphorylation during smooth muscle contractions.
Article in Analytical biochemistry, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed, 10 citations in OpenAlex.
- Antibody-oligonucleotide conjugates for spatial proteomics: principles, applications, and challenges.Acta biochimica et biophysica Sinica · 2025Article
- Mfge8 is expressed by pericytes in gastric antrum submucosa from patients with obesity.American journal of physiology. Cell physiology · 2023Article
- Article
- Molecular probes for cellular imaging of post-translational proteoforms.RSC chemical biology · 2022Review
- Mfge8 attenuates human gastric antrum smooth muscle contractions.Journal of muscle research and cell motility · 2021Article
- Analyzing the Integrin Adhesome by In Situ Proximity Ligation Assay.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Increased talin-vinculin spatial proximities in livers in response to spotted fever group rickettsial and Ebola virus infections.Laboratory investigation; a journal of technical methods and pathology · 2020Article
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
Antibody-based in situ proximity ligation assays (isPLA) have the potential to study protein phosphorylation and protein interactions with spatial resolution in intact tissues. However, the application of isPLA at the tissue level is limited by a lack of appropriate positive and negative controls and the difficulty in accounting for changes in tissue shape. Here we demonstrate a set of experimental and computational approaches using gastric fundus smooth muscles to improve the validity of quantitative isPLA. Appropriate positive and negative biological controls and PLA technical controls were selected to ensure experimental rigor. To account for changes in morphology between relaxed and contracted smooth muscles, target PLA spots were normalized to smooth muscle myosin light chain 20 PLA spots or the cellular cross-sectional areas. We describe the computational steps necessary to filter out false-positive improperly sized spots and set the thresholds for counting true positive PLA spots to quantify the PLA signals. We tested our approach by examining protein phosphorylation and protein interactions in smooth muscle myofilament Ca
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Registered trials
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