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STX3, a known regulator of apical trafficking in epithelial cells, was previously understood to be absent in human RPE cells, with its functions thought to be compensated by STX1A. However, our results on SNARE mRNA expression profile in RPE detected the presence of 2 splice variants of STX3. Further investigation in donor retina, primary hRPE, and ARPE\u201019 cells revealed detectable levels of STX3 mRNA and protein. STX3 knockdown in ARPE19 resulted in a significant reduction of tight junction (TJ) proteins, compromising TJ assembly, highlighting the critical role of STX3 in maintaining RPE integrity. In addition, immunoprecipitation followed by LC\u2010MS\/MS analysis revealed that STX3 and STX1A have a distinct novel protein interactome in RPE. This study identified unique and shared interactants for STX3 and STX1A, suggesting a broader role for RPE beyond its traditional photoreceptor support function. This further emphasizes the biological significance of STX1A and STX3 in maintaining retinal homeostasis, which could facilitate the development of novel therapeutic strategies for retinal disorders.<\/jats:p>","DOI":"10.1002\/cbf.70294","type":"journal-article","created":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T09:46:46Z","timestamp":1788515206000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Syntaxin\u20103 Regulates Tight Junction Assembly in Human Retinal Pigment Epithelium"],"prefix":"10.1002","volume":"44","author":[{"given":"Athira","family":"Ramesh","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Molecular Biology Pondicherry University Puducherry India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9778-5400","authenticated-orcid":false,"given":"Suganya","family":"Sivagurunathan","sequence":"additional","affiliation":[{"name":"RS Mehta Jain Department of Biochemistry and Cell Biology Vision Research Foundation Chennai India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Navya Valavath","family":"Baburajan","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology Pondicherry University Puducherry India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3180-4429","authenticated-orcid":false,"given":"Subbulakshmi","family":"Chidambaram","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology Pondicherry University Puducherry India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,4]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.preteyeres.2014.06.002"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00021.2004"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.exer.2014.04.021"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.94.24.12932"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ceb.2019.08.001"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1034\/j.1600-0854.2001.21202.x"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.3389\/fnana.2022.983151"},{"key":"e_1_2_10_9_1","doi-asserted-by":"crossref","unstructured":"X.Li H.Low M.Miura andT.Weimbs(2002). 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