Optimizing In Situ Proximity Ligation Assays for Mitochondria, ER, or MERC Markers in Skeletal Muscle Tissue and Cells
Proximity ligation assays (PLAs) use specific antibodies to detect endogenous protein-protein interactions. PLAs are a highly useful biochemical technique that allow two proteins within proximity to be visualized with fluorescent probes amplified by PCR. While this technique has gained prominence, the use of a PLA in mouse skeletal muscle (SkM) is novel. In this article, we discuss how the PLA method can be used in SkM to study the protein-protein interactions within mitochondria-endoplasmic reticulum contact sites (MERCs). © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Proximity ligation assay for skeletal muscle tissue and myoblast for MERC proteins.
Authors
- Antentor Hinton (ORCID: https://orcid.org/0000-0002-7730-952X)
- Andrea G. Marshall (ORCID: https://orcid.org/0000-0001-9997-4478)
- Larry Vang (ORCID: https://orcid.org/0000-0001-9098-3650)
- Neng Vue
- Edgar Garza-López (ORCID: https://orcid.org/0000-0003-0243-0816)
- Jennifer Angeline Gaddy (ORCID: https://orcid.org/0000-0002-2192-4224)
- Steven M. Damo (ORCID: https://orcid.org/0000-0002-1483-7712)
- Sandra Murray (ORCID: https://orcid.org/0000-0002-8267-9138)
- Celestine N. Wanjalla (ORCID: https://orcid.org/0000-0001-9159-5414)
- Blanca Rodríguez (ORCID: https://orcid.org/0000-0001-6361-3339)
- Johnathan Moore (ORCID: https://orcid.org/0000-0002-3207-5360)
- Kit Neikirk (ORCID: https://orcid.org/0000-0003-0994-0333)
- Heather K. Beasley (ORCID: https://orcid.org/0000-0002-4636-2889)
- Margaret Mungai (ORCID: https://orcid.org/0000-0003-3292-1552)
- Zer Vue (ORCID: https://orcid.org/0000-0002-3502-8906)
- Desiree Nguyen
- Jianqiang Shao
- Amber Crabtree (ORCID: https://orcid.org/0000-0003-1156-1658)
- Jamaine Davis (ORCID: https://orcid.org/0000-0002-5078-1306)
- Izabella Marie Rabago
- Dominique C. Stephens
- Kyrin Turner
- Chanel Harris
- Ashton Oliver (ORCID: https://orcid.org/0009-0001-9874-2621)
- Han Le
- Andy Barillas
Institutions
- Meharry Medical College (US)
- University of Iowa (US)
- United States Department of Veterans Affairs (US)
- University of Pittsburgh (US)
- Vanderbilt University (US)
- VA Tennessee Valley Healthcare System (US)
- Vanderbilt University Medical Center (US)
- Fisk University (US)
Publication Details
- Journal
- Current Protocols
- Published
- 2025-02-01
- DOI
- https://doi.org/10.1002/cpz1.70043
- Primary Topic
- Mitochondrial Function and Pathology
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- National Science Foundation
- U.S. Department of Veterans Affairs
- Silicon Valley Community Foundation
- Bristol-Myers Squibb
- Vanderbilt University
- National Institutes of Health