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The Src-family kinase Lyn plays activating and inhibitory roles downstream of TLRs, yet distinct functions of the Lyn splice variants LynA and LynB in TLR signaling had not been investigated. We used isoform-specific Lyn knockout mice (LynA\n                  <jats:sup>KO<\/jats:sup>\n                  and LynB\n                  <jats:sup>KO<\/jats:sup>\n                  ) to interrogate the contribution of each isoform to TLR signaling in bone marrow-derived macrophages. Bulk RNA sequencing and cytokine analyses revealed that complete Lyn deficiency (Lyn\n                  <jats:sup>KO<\/jats:sup>\n                  ) dampens TLR4- and TLR7-induced inflammatory gene expression and TNF production, but enhances the expression of genes responsible for synthesizing the extracellular matrix and promoting proliferation. Despite a reduction in total Lyn levels, the expression of either LynA or LynB alone was sufficient to preserve wild-type transcriptional responses and TNF production in response to the TLR7 agonist R848. However, LyA\n                  <jats:sup>KO<\/jats:sup>\n                  and LynB\n                  <jats:sup>KO<\/jats:sup>\n                  macrophages did have partially impaired TNF production in response to the TLR4 agonist lipopolysaccharide. Additionally, LynA\n                  <jats:sup>KO<\/jats:sup>\n                  and LynB\n                  <jats:sup>KO<\/jats:sup>\n                  macrophages were hyperproliferative, like Lyn\n                  <jats:sup>KO<\/jats:sup>\n                  cells. These data suggest that Lyn promotes macrophage activation downstream of TLRs and restrains aberrant proliferation and matrix deposition in a dose-dependent rather than isoform-specific manner.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Summary Sentence<\/jats:title>\n                  <jats:p>RNA sequencing and functional assays demonstrate that both LynA and LynB restrict macrophage proliferation and drive TLR-induced ECM-remodeling and inflammatory cytokine production.<\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2025.07.07.663098","type":"posted-content","created":{"date-parts":[[2025,7,11]],"date-time":"2025-07-11T01:45:17Z","timestamp":1752198317000},"source":"Crossref","is-referenced-by-count":0,"title":["Total, not isoform-specific, Lyn expression by macrophages promotes TLR activation and restricts proliferation"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1988-5163","authenticated-orcid":false,"given":"Anders J.","family":"Lindstedt","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3482-0958","authenticated-orcid":false,"given":"Joseph T.","family":"Greene","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5168-5829","authenticated-orcid":false,"given":"Tanya S.","family":"Freedman","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"issue":"1","key":"2025071322150517000_2025.07.07.663098v1.1","first-page":"e03005","article-title":"CARD9 Is Required for Classical Macrophage Activation and the Induction of Protective Immunity against Pulmonary Cryptococcosis","volume":"11","year":"2020","journal-title":"mBio"},{"key":"2025071322150517000_2025.07.07.663098v1.2","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1102154"},{"key":"2025071322150517000_2025.07.07.663098v1.3","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20100086"},{"key":"2025071322150517000_2025.07.07.663098v1.4","doi-asserted-by":"publisher","DOI":"10.1183\/13993003.02441-2018"},{"key":"2025071322150517000_2025.07.07.663098v1.5","doi-asserted-by":"publisher","DOI":"10.1164\/rccm.201010-1719OC"},{"key":"2025071322150517000_2025.07.07.663098v1.6","doi-asserted-by":"publisher","DOI":"10.1007\/s10753-014-9941-y"},{"issue":"2","key":"2025071322150517000_2025.07.07.663098v1.7","first-page":"80","article-title":"Macrophage Checkpoint Blockade: Results from Initial Clinical Trials, Binding Analyses, and CD47-SIRP\u03b1 Structure-Function","volume":"3","year":"2020","journal-title":"Antib Ther"},{"key":"2025071322150517000_2025.07.07.663098v1.8","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0076147"},{"key":"2025071322150517000_2025.07.07.663098v1.9","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-019-1456-0"},{"issue":"1","key":"2025071322150517000_2025.07.07.663098v1.10","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/j.cels.2017.06.014","article-title":"Systematic Investigation of Multi-TLR Sensing Identifies Regulators of Sustained Gene Activation in Macrophages","volume":"5","year":"2017","journal-title":"Cell Syst"},{"key":"2025071322150517000_2025.07.07.663098v1.11","doi-asserted-by":"publisher","DOI":"10.1038\/ni1280"},{"key":"2025071322150517000_2025.07.07.663098v1.12","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.180.6.3729"},{"key":"2025071322150517000_2025.07.07.663098v1.13","doi-asserted-by":"publisher","DOI":"10.1038\/ni1028"},{"key":"2025071322150517000_2025.07.07.663098v1.14","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M306793200"},{"key":"2025071322150517000_2025.07.07.663098v1.15","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M111.266528"},{"key":"2025071322150517000_2025.07.07.663098v1.16","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2011.05.006"},{"key":"2025071322150517000_2025.07.07.663098v1.17","doi-asserted-by":"publisher","DOI":"10.1016\/j.cyto.2009.08.010"},{"key":"2025071322150517000_2025.07.07.663098v1.18","doi-asserted-by":"publisher","DOI":"10.1016\/j.febslet.2006.06.100"},{"key":"2025071322150517000_2025.07.07.663098v1.19","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajpath.2012.09.007"},{"key":"2025071322150517000_2025.07.07.663098v1.20","doi-asserted-by":"publisher","DOI":"10.1038\/nri1391"},{"key":"2025071322150517000_2025.07.07.663098v1.21","doi-asserted-by":"crossref","unstructured":"Duan, T. ; Du, Y. ; Xing, C. ; Wang, H. 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