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Transforming growth factor-\u03b2 (TGF-\u03b2) is the most powerful known driver of myofibroblast differentiation. TGF-\u03b2 signals through transmembrane receptor serine\/threonine kinases that phosphorylate Smad transcription factors (Smad2\/3) leading to activation of transcription of target genes. Heterotrimeric G proteins mediate distinct signaling from seven-transmembrane G protein coupled receptors, which are not known to be linked to Smad activation. We tested whether G protein signaling plays any role in TGF-\u03b2-induced myofibroblast differentiation, using primary cultured human lung fibroblasts. Activation of G\u03b1s by cholera toxin blocked TGF-\u03b2-induced myofibroblast differentiation without affecting Smad2\/3 phosphorylation. Neither inhibition of G\u03b1i by pertussis toxin nor siRNA-mediated combined knockdown of G\u03b1q and G\u03b111 had a significant effect on TGF-\u03b2-induced myofibroblast differentiation. In contrast, combined knockdown of G\u03b112 and G\u03b113 significantly inhibited TGF-\u03b2-stimulated expression of myofibroblast marker proteins (collagen-1, fibronectin, smooth-muscle \u03b1-actin), with siG\u03b112 being significantly more potent than siG\u03b113. Mechanistically, combined knockdown of G\u03b112 and G\u03b113 resulted in substantially reduced phosphorylation of Smad2 and Smad3 in response to TGF-\u03b2, which was accompanied by a significant decrease in the expression of TGF-\u03b2 receptors (TGFBR1, TGFBR2) and of Smad3. Thus, our study uncovers a novel role of G\u03b112\/13 proteins in the control of TGF-\u03b2 signaling and myofibroblast differentiation.<\/jats:p>","DOI":"10.1042\/bcj20240317","type":"journal-article","created":{"date-parts":[[2024,12,2]],"date-time":"2024-12-02T17:17:06Z","timestamp":1733159826000},"page":"1937-1948","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["G\u03b112 and G\u03b113 proteins are required for transforming growth factor-\u03b2-induced myofibroblast differentiation"],"prefix":"10.1042","volume":"481","author":[{"given":"Eleanor B.","family":"Reed","sequence":"first","affiliation":[{"name":"1Department of Medicine, Section of Pulmonary and Critical Care Medicine, The University of Chicago, Chicago, IL, U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Albert","family":"Sitikov","sequence":"additional","affiliation":[{"name":"1Department of Medicine, Section of Pulmonary and Critical Care Medicine, The University of Chicago, Chicago, IL, U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kun Woo D.","family":"Shin","sequence":"additional","affiliation":[{"name":"1Department of Medicine, Section of Pulmonary and Critical Care Medicine, The University of Chicago, Chicago, IL, U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Robert B.","family":"Hamanaka","sequence":"additional","affiliation":[{"name":"1Department of Medicine, Section of Pulmonary and Critical Care Medicine, The University of Chicago, Chicago, IL, U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Reng\u00fcl","family":"Cetin-Atalay","sequence":"additional","affiliation":[{"name":"1Department of Medicine, Section of Pulmonary and Critical Care Medicine, The University of Chicago, Chicago, IL, U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"G\u00f6khan M.","family":"Mutlu","sequence":"additional","affiliation":[{"name":"1Department of Medicine, Section of Pulmonary and Critical Care Medicine, The University of Chicago, Chicago, IL, U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexander A.","family":"Mongin","sequence":"additional","affiliation":[{"name":"2Department of Neuroscience and Experimental Therapeutics, Albany Medical College, Albany, NY, U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6096-5822","authenticated-orcid":false,"given":"Nickolai O.","family":"Dulin","sequence":"additional","affiliation":[{"name":"1Department of Medicine, Section of Pulmonary and Critical Care Medicine, The University of Chicago, Chicago, IL, U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"288","published-online":{"date-parts":[[2024,12,13]]},"reference":[{"key":"2024121313354754700_BCJ-2024-0317C1","first-page":"4776","article-title":"Synergistic interaction of two classes of transforming growth factors from murine sarcoma cells","volume":"42","year":"1982","journal-title":"Cancer Res."},{"key":"2024121313354754700_BCJ-2024-0317C2","doi-asserted-by":"publisher","first-page":"6264","DOI":"10.1073\/pnas.80.20.6264","article-title":"Sarcoma growth factor from conditioned medium of virally transformed cells is composed of both type alpha and type beta transforming growth factors","volume":"80","year":"1983","journal-title":"Proc. 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