{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T18:10:12Z","timestamp":1790014212318,"version":"4.0.1"},"reference-count":37,"publisher":"Public Library of Science (PLoS)","issue":"9","license":[{"start":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T00:00:00Z","timestamp":1789948800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>Verteporfin (VP) is widely used as a light-independent pharmacologic inhibitor of the YAP-TEAD transcriptional complex, despite its potent photosensitizing properties. Here we show that VP-associated high\u2013molecular\u2013weight complexes and apparent depletion of key proteins on immunoblots can be substantially driven by ambient-light exposure during post-lysis handling, more than by light during cellular treatment. In this study, the hypothesis is tested independently using MEL270, HEK293, and MCF-7 cells treated with therapeutic and supra-therapeutic concentrations of VP under four systematically varied light\/dark conditions. The results demonstrate that high-molecular-weight complex (HMWC) formation for p62, DIAP1, ROCK1, YAP, and phospho-YAP is driven principally by light exposure during sample processing rather than during cell treatment, occurs in cell-free lysates within 1 hour of ambient-light exposure, and is markedly attenuated by both N-acetylcysteine and L-histidine, consistent with contributions from both radical-mediated and singlet oxygen-mediated photochemistry. Notably, YAP and phospho-YAP are highly susceptible to photo-cross-linking, whereas transcription enhancer factor 1 (TEF1)\/TEAD1 remains comparatively resistant, a pattern that mimics selective YAP-TEAD inhibition on standard immunoblots. In a single-experiment CCK-8 assay, viability was markedly lower under light than in darkness at 24 hours, an observation consistent with the immunoblot findings but requiring independent replication. These findings support the light-artifact hypothesis and may have implications for how preclinical data on VP are interpreted in the design of clinical investigations.<\/jats:p>","DOI":"10.1371\/journal.pone.0358654","type":"journal-article","created":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T17:39:18Z","timestamp":1790012358000},"page":"e0358654","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Ambient light drives verteporfin-induced protein cross-linking during standard laboratory sample processing"],"prefix":"10.1371","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-0766-2992","authenticated-orcid":true,"given":"Konstantinos G.","family":"Baroutis","sequence":"first","affiliation":[{"name":"Retina Service, Ines and Frederick Yeatts Lab in Retina Research","place":["Massachusetts, Boston, Massachusetts, United States of America"],"department":["Department of Ophthalmology"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Victor San Martin C.","family":"Corr\u00eaa","sequence":"additional","affiliation":[{"name":"Retina Service, Ines and Frederick Yeatts Lab in Retina Research","place":["Massachusetts, Boston, Massachusetts, United States of America"],"department":["Department of Ophthalmology"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0524-4752","authenticated-orcid":true,"given":"Dimitrios","family":"Ntentakis","sequence":"additional","affiliation":[{"name":"Retina Service, Ines and Frederick Yeatts Lab in Retina Research","place":["Massachusetts, Boston, Massachusetts, United States of America"],"department":["Department of Ophthalmology"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2844-1046","authenticated-orcid":true,"given":"Toshio","family":"Narimatsu","sequence":"additional","affiliation":[{"name":"Retina Service, Ines and Frederick Yeatts Lab in Retina Research","place":["Massachusetts, Boston, Massachusetts, United States of America"],"department":["Department of Ophthalmology"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Evangelos S.","family":"Gragoudas","sequence":"additional","affiliation":[{"name":"Retina Service, Ines and Frederick Yeatts Lab in Retina Research","place":["Massachusetts, Boston, Massachusetts, United States of America"],"department":["Department of Ophthalmology"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joan W.","family":"Miller","sequence":"additional","affiliation":[{"name":"Retina Service, Ines and Frederick Yeatts Lab in Retina Research","place":["Massachusetts, Boston, Massachusetts, United States of America"],"department":["Department of Ophthalmology"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lucy H.","family":"Young","sequence":"additional","affiliation":[{"name":"Retina Service, Ines and Frederick Yeatts Lab in Retina Research","place":["Massachusetts, Boston, Massachusetts, United States of America"],"department":["Department of Ophthalmology"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8622-6478","authenticated-orcid":true,"given":"Demetrios G.","family":"Vavvas","sequence":"additional","affiliation":[{"name":"Retina Service, Ines and Frederick Yeatts Lab in Retina Research","place":["Massachusetts, Boston, Massachusetts, United States of America"],"department":["Department of Ophthalmology"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,9,21]]},"reference":[{"issue":"6","key":"pone.0358654.ref001","doi-asserted-by":"crossref","first-page":"901","DOI":"10.1080\/09553008514553131","article-title":"Photosensitization with derivatives of haematoporphyrin","volume":"49","author":"D Kessel","year":"1986","journal-title":"Int J Radiat Biol Relat Stud Phys Chem Med"},{"key":"pone.0358654.ref002","doi-asserted-by":"crossref","unstructured":"Photodynamic therapy of subfoveal choroidal neovascularization in age-related macular degeneration with verteporfin: one-year results of 2 randomized clinical trials--TAP report. 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