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Macrophages are the cell type preferentially infected by <jats:italic>T. gondii <\/jats:italic>in vivo. Protein phosphorylation is an important posttranslational modification involved in diverse cellular functions. A rapidly accelerated fibrosarcoma kinase (A-Raf) is a member of the Raf family of serine\/threonine protein kinases that is necessary for MAPK activation. Our previous research found that knockout of A-Raf could reduce <jats:italic>T. gondii<\/jats:italic>-induced apoptosis in porcine alveolar macrophages (3D4\/21 cells). However, limited information is available on protein phosphorylation variations and the role of A-Raf in macrophages infected with <jats:italic>T. gondii<\/jats:italic>.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Methods<\/jats:title>\n                <jats:p>We used immobilized metal affinity chromatography (IMAC) in combination with liquid chromatography tandem mass spectrometry (LC\u2013MS\/MS) to profile changes in phosphorylation in <jats:italic>T. gondii-<\/jats:italic>infected 3D4\/21 and 3D4\/21-\u0394<jats:italic>Araf<\/jats:italic> cells.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>A total of 1647 differentially expressed phosphorylated proteins (DEPPs) with 3876 differentially phosphorylated sites (DPSs) were identified in <jats:italic>T. gondii<\/jats:italic>-infected 3D4\/21 cells (p3T group) when compared with uninfected 3D4\/21 cells (pho3 group), and 959 DEPPs with 1540 DPSs were identified in the p3T group compared with infected 3D4\/21-\u0394<jats:italic>Araf<\/jats:italic> cells (p3KT group). Venn analysis revealed 552 DPSs corresponding to 406 DEPPs with the same phosphorylated sites when comparing p3T\/pho3 versus p3T\/p3KT, which were identified as DPSs and DEPPs that were directly or indirectly related to A-Raf.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Conclusions<\/jats:title>\n                <jats:p>Our results revealed distinct responses of macrophages to <jats:italic>T. gondii<\/jats:italic> infection and the potential roles of A-Raf in fighting infection via phosphorylation of crucial proteins.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Graphical Abstract<\/jats:title>\n                \n              <\/jats:sec>","DOI":"10.1186\/s13071-024-06273-x","type":"journal-article","created":{"date-parts":[[2024,4,20]],"date-time":"2024-04-20T10:02:01Z","timestamp":1713607321000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Phosphoproteomic analysis reveals changes in A-Raf-related protein phosphorylation in response to Toxoplasma gondii infection in porcine macrophages"],"prefix":"10.1186","volume":"17","author":[{"given":"Dingzeyang","family":"Su","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shifan","family":"Zhu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kangzhi","family":"Xu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhaofeng","family":"Hou","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fuxing","family":"Hao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fan","family":"Xu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yifan","family":"Lin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuyang","family":"Zhu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dandan","family":"Liu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qiangde","family":"Duan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xinjun","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuguo","family":"Yuan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jinjun","family":"Xu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jianping","family":"Tao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2024,4,20]]},"reference":[{"key":"6273_CR1","doi-asserted-by":"publisher","first-page":"1375","DOI":"10.1017\/S0031182012000765","volume":"139","author":"JP Dubey","year":"2012","unstructured":"Dubey JP, Lago EG, Gennari SM, Su C, Jones JL. 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