ArticlePloS one2015
Immuno-Northern Blotting: Detection of RNA Modifications by Using Antibodies against Modified Nucleosides.
Article in PloS one, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 44 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
44 citing papers in PubMed, 85 citations in OpenAlex.
- The crucial role of m6A methylation modification in nasopharyngeal carcinoma.Discover oncology · 2026Review
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- Advances in Quantitative Techniques for Mapping RNA Modifications.Life (Basel, Switzerland) · 2025Review
- The role of NTAG. Theoretical and applied genetics. Theoretische und angewandte Genetik · 2025Review
- Small nucleolar RNAs promote the restoration of muscle differentiation defects in cells from myotonic dystrophy type 1.Nucleic acids research · 2025Article
- Detection, molecular function and mechanisms of m5C in cancer.Clinical and translational medicine · 2025Review
- RNA modifications and their role in gene expression.Frontiers in molecular biosciences · 2025Review
- Biological functions of 5-methylcytosine RNA-binding proteins and their potential mechanisms in human cancers.Frontiers in oncology · 2025Review
- Navigating the pitfalls of mapping DNA and RNA modifications.Nature reviews. Genetics · 2023Review
- RNA methylation in plants: An overview.Frontiers in plant science · 2023Review
- Analysis of NBio-protocol · 2022Article
- Epitranscriptomic Modifications Modulate Normal and Pathological Functions in CNS.Translational stroke research · 2022Review
- Codon-Reduced Protein Synthesis With Manipulating tRNA Components in Cell-Free System.Frontiers in bioengineering and biotechnology · 2022Review
- The emerging therapeutic target of dynamic and reversible N6-methyladenosine modification during cancer development.Frontiers in oncology · 2022Review
- Advances in mRNA 5-methylcytosine modifications: Detection, effectors, biological functions, and clinical relevance.Molecular therapy. Nucleic acids · 2021Review
- Targeting the RNA demethylase FTO for cancer therapy.RSC chemical biology · 2021Review
- m6A RNA Methylation in Systemic Autoimmune Diseases-A New Target for Epigenetic-Based Therapy?Pharmaceuticals (Basel, Switzerland) · 2021Review
- Analysis of RNA Modifications by Second- and Third-Generation Deep Sequencing: 2020 Update.Genes · 2021Review
- The role of mMolecular cancer · 2021Review
- m6A Modification in Non-Coding RNA: The Role in Cancer Drug Resistance.Frontiers in oncology · 2021Review
Corrections and comments
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Authors and funding
14 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The biological roles of RNA modifications are still largely not understood. Thus, developing a method for detecting RNA modifications is important for further clarification. We developed a method for detecting RNA modifications called immuno-northern blotting (INB) analysis and herein introduce its various capabilities. This method involves the separation of RNAs using either polyacrylamide or agarose gel electrophoresis, followed by transfer onto a nylon membrane and subsequent immunoblotting using antibodies against modified nucleosides for the detection of specific modifications. We confirmed that INB with the antibodies for 1-methyladenosine (m1A), N6-methyladenosine (m6A), pseudouridine, and 5-methylcytidine (m5C) showed different modifications in a variety of RNAs from various species and organelles. INB with the anti-m5C antibody revealed that the antibody cross-reacted with another modification on DNA, suggesting the application of this method for characterization of the antibody for modified nucleosides. Additionally, using INB with the antibody for m1A, which is a highly specific modification in eukaryotic tRNA, we detected tRNA-derived fragments known as tiRNAs under the cellular stress response, suggesting the application for tracking target RNA containing specific modifications. INB with the anti-m6A antibody confirmed the demethylation of m6A by the specific demethylases fat mass and obesity-associated protein (FTO) and ALKBH5, suggesting its application for quantifying target modifications in separated RNAs. Furthermore, INB demonstrated that the knockdown of FTO and ALKBH5 increased the m6A modification in small RNAs as well as in mRNA. The INB method has high specificity, sensitivity, and quantitative capability, and it can be employed with conventional experimental apparatus. Therefore, this method would be useful for research on RNA modifications and metabolism.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.