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However, although much information has been gleaned from FRET studies on cardiomyocytes from larger species, experiments on adult cardiomyocytes from mice have been difficult at best. Thus the large variety of genetic mouse models cannot be easily used for this type of study. Here we develop cell culture conditions for adult mouse cardiomyocytes that permit robust expression of adenoviral FRET biosensors and reproducible FRET experimentation. We find that addition of 6.25 \u00b5M blebbistatin or 20 \u00b5M (S)-nitro-blebbistatin to a minimal essential medium containing 10 mM HEPES and 0.2% BSA maintains morphology of cardiomyocytes from physiological, pathological, and transgenic mouse models for up to 50 h after adenoviral infection. This provides a 10\u201315-h time window to perform reproducible FRET readings using a variety of CFP\/YFP sensors between 30 and 50 h postinfection. The culture is applicable to cardiomyocytes isolated from transgenic mouse models as well as models with cardiac diseases. Therefore, this study helps scientists to disentangle complicated signaling networks important in health and disease of cardiomyocytes.<\/jats:p>","DOI":"10.1085\/jgp.201812119","type":"journal-article","created":{"date-parts":[[2018,9,21]],"date-time":"2018-09-21T09:37:37Z","timestamp":1537522657000},"page":"1567-1582","update-policy":"https:\/\/doi.org\/10.1085\/jgp.crossmarkpolicy","source":"Crossref","is-referenced-by-count":24,"title":["Illuminating cell signaling with genetically encoded FRET biosensors in adult mouse cardiomyocytes"],"prefix":"10.1085","volume":"150","author":[{"given":"Gopireddy Raghavender","family":"Reddy","sequence":"first","affiliation":[{"name":"Department of Pharmacology, University of California at Davis, Davis, CA 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7665-1860","authenticated-orcid":true,"given":"Toni M.","family":"West","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of California at Davis, Davis, CA 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhong","family":"Jian","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of California at Davis, Davis, CA 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8916-7533","authenticated-orcid":true,"given":"Mark","family":"Jaradeh","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of California at Davis, Davis, CA 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qian","family":"Shi","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of California at Davis, Davis, CA 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of California at Davis, Davis, CA 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4818-9570","authenticated-orcid":true,"given":"Ye","family":"Chen-Izu","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of California at Davis, Davis, CA 1"},{"name":"Department of Bioengineering, University of California at Davis, Davis, CA 2"},{"name":"Department of Internal Medicine\/Cardiology, University of California at Davis, Davis, CA 3"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1786-9143","authenticated-orcid":true,"given":"Yang K.","family":"Xiang","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of California at Davis, Davis, CA 1"},{"name":"Veterans Affairs Northern California Health Care System, Mather, CA 4"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"291","published-online":{"date-parts":[[2018,9,21]]},"reference":[{"key":"2026041423520402100_bib1","doi-asserted-by":"publisher","first-page":"909","DOI":"10.1161\/CIRCRESAHA.116.309202","article-title":"A simplified, Langendorff-free method for concomitant isolation of viable cardiac myocytes and nonmyocytes from the adult mouse heart: novelty and significance","volume":"119","author":"Ackers-Johnson","year":"2016","journal-title":"Circ. 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