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To enable direct analysis of native assemblies, we generated a knock-in mouse in which the endogenous \u03b3 subunit is fused at its C terminus to mVenus, a 3C protease cleavage site, and a 3xFLAG epitope (ENaC\u03b3-VF). The tag preserves physiological ENaC function, as ENaC\u03b3-VF mice display normal electrolyte handling, benzamil affinity, and amiloride-sensitive Na\u207a:K\u207a responses indistinguishable from wild-type animals. Using fluorescence-detection size-exclusion chromatography and single-molecule pull-down, we directly monitor intact native ENaC complexes from lung, kidney, and colon and uncover marked tissue-to-tissue differences in channel abundance and apparent complex size. Dual-color analysis with a fluorescent Fab against ENaC\u03b1 marks fully assembled \u03b1\u03b2\u03b3 channels, while \u03b3-based fluorescence reports the broader population of \u03b3-containing assemblies. In combination, the ENaC\u03b3-VF line provides a biochemical anchor for identifying regulatory and trafficking proteins that co-purify with native ENaC complexes. These data show that ENaC architecture\n                  <jats:italic>in vivo<\/jats:italic>\n                  is heterogeneous, and establish ENaC\u03b3-VF mice as a platform for dissecting how epithelial environments shape ENaC assembly, composition, and regulation.\n                <\/jats:p>","DOI":"10.64898\/2026.01.23.701393","type":"posted-content","created":{"date-parts":[[2026,1,26]],"date-time":"2026-01-26T04:30:10Z","timestamp":1769401810000},"source":"Crossref","is-referenced-by-count":0,"title":["Differential Assembly of Native ENaC Complexes Across Mouse Epithelial Tissues"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3867-7610","authenticated-orcid":false,"given":"Arpita","family":"Bharadwaj","sequence":"first","affiliation":[{"name":"Vollum Institute, Oregon Health & Science University, Portland, OR 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