ArticlePloS one2014
Sensitivity and specificity of in situ proximity ligation for protein interaction analysis in a model of steatohepatitis with Mallory-Denk bodies.
Article in PloS one, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed.
- Proximity Ligation Assay: From a Foundational Principle to a Versatile Platform for Molecular and Translational Research.Biomolecules · 2025Review
- Tools to investigate the cell surface: Proximity as a central concept in glycoRNA biology.Cell chemical biology · 2024Review
- The formin DAAM1 regulates the deubiquitinase activity of USP10 and integrin homeostasis.European journal of cell biology · 2023Article
- Candida albicans stimulates formation of a multi-receptor complex that mediates epithelial cell invasion during oropharyngeal infection.PLoS pathogens · 2023Article
- ABRAXAS1 orchestrates BRCA1 activities to counter genome destabilizing repair pathways-lessons from breast cancer patients.Cell death & disease · 2023Article
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- Genome Editing and Fatty Liver.Advances in experimental medicine and biology · 2023Review
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- Activation of EphA2-EGFR signaling in oral epithelial cells by Candida albicans virulence factors.PLoS pathogens · 2021Article
- A Novel Mechanism Underlying Antiviral Activity of an Influenza Virus M2-Specific Antibody.Journal of virology · 2020Article
- Modeling Non-Alcoholic Fatty Liver Disease (NAFLD) Using "Good-Fit" Genome-Editing Tools.Cells · 2020Review
- Activation of β1 integrins and caveolin-1 by TF/FVIIa promotes IGF-1R signaling and cell survival.Apoptosis : an international journal on programmed cell death · 2020Article
- The dynamic emergence of GATA1 complexes identified inHaematologica · 2020Article
- Starvation-induced autophagy via calcium-dependent TFEB dephosphorylation is suppressed by Shigyakusan.PloS one · 2020Article
- EGF-activated PI3K/Akt signalling coordinates leucine uptake by regulating LAT3 expression in prostate cancer.Cell communication and signaling : CCS · 2019Article
- Decreased synthesis of ribosomal proteins in tauopathy revealed by non-canonical amino acid labelling.The EMBO journal · 2019Article
- Human papillomavirus type 16 E6 and E7 oncoproteins interact with the nuclear p53-binding protein 1 in an in vitro reconstructed 3D epithelium: new insights for the virus-induced DNA damage response.Virology journal · 2018Article
- The BH3-only protein BAD mediates TNFα cytotoxicity despite concurrent activation of IKK and NF-κB in septic shock.Cell research · 2018Article
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The in situ proximity ligation assay (isPLA) is an increasingly used technology for in situ detection of protein interactions, post-translational modifications, and spatial relationships of antigens in cells and tissues, in general. In order to test its performance we compared isPLA with immunofluorescence microscopy by analyzing protein interactions in cytoplasmic protein aggregates, so-called Mallory Denk bodies (MDBs). These structures represent protein inclusions in hepatocytes typically found in human steatohepatitis and they can be generated in mice by feeding of 3,5-diethoxy-carbonyl-1,4-dihydrocollidine (DDC). We investigated the colocalization of all three key MDB components, namely keratin 8 (K8), keratin 18 (K18), and p62 (sequestosome 1) by isPLA and immunofluorescence microscopy. Sensitivity and specificity of isPLA was assessed by using Krt8-/- and Krt18-/- mice as biological controls, along with a series of technical controls. isPLA signal visualization is a robust technology with excellent sensitivity and specificity. The biological relevance of signals generated critically depends on the performance of antibodies used, which requires careful testing of antibodies like in immunofluorescence microscopy. There is a clear advantage of isPLA in visualizing protein co-localization, particularly when antigens are present at markedly different concentrations. Furthermore, isPLA is superior to confocal microscopy with respect to spatial resolution of colocalizing antigens. Disadvantages compared to immunofluorescence are increased costs and longer duration of the laboratory protocol.
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