{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,24]],"date-time":"2026-07-24T01:57:09Z","timestamp":1784858229474,"version":"3.55.0"},"posted":{"date-parts":[[2023,10,14]]},"group-title":"Cell Biology","reference-count":23,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,10,14]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  The plasma membrane is a well-organized structure of lipids and proteins, segmented into lipid compartments under 200 nm in size. This specific spatial patterning is crucial for the function of proteins and necessitates super-resolution imaging for its elucidation. Here, we establish that the genetically encoded enhanced green fluorescent protein (EGFP), when combined with direct optical reconstruction microscopy (dSTORM), tracks shear- and cholesterol-induced nanoscopic patterning of potassium channels overexpressed in HEK293T cells. Leveraging EGFP in dSTORM (EGFP-STORM), our findings indicate that cholesterol directs the C-terminus of TWIK-related potassium channel (TREK-1) to ceramide-enriched lipid ganglioside (GM1) clusters. In the absence of the C-terminus, the channel associates with phosphatidylinositol 4,5-bisphosphate (PIP\n                  <jats:sub>2<\/jats:sub>\n                  ) cluster. Similarly, cholesterol derived from astrocytes repositions EGFP-tagged inward-rectifying potassium (Kir) channels into GM1 clusters. Without cholesterol, the channel aligns with PIP\n                  <jats:sub>2<\/jats:sub>\n                  lipids. We deduce that cholesterol\u2019s interaction with Kir sequesters the channel, separating it from its activating lipid PIP\n                  <jats:sub>2<\/jats:sub>\n                  . Fundamentally, a genetically encoded EGFP tag should make any protein amenable to dSTORM analysis.\n                <\/jats:p>","DOI":"10.1101\/2023.10.13.561998","type":"posted-content","created":{"date-parts":[[2023,10,14]],"date-time":"2023-10-14T16:05:10Z","timestamp":1697299510000},"source":"Crossref","is-referenced-by-count":8,"title":["Super-resolution imaging of potassium channels with genetically encoded EGFP"],"prefix":"10.64898","author":[{"given":"Isabelle M.","family":"Call","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julian L.","family":"Bois","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Scott B.","family":"Hansen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2023101823050710000_2023.10.13.561998v2.1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.200802376"},{"key":"2023101823050710000_2023.10.13.561998v2.2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2004259117"},{"key":"2023101823050710000_2023.10.13.561998v2.3","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1202"},{"key":"2023101823050710000_2023.10.13.561998v2.4","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms13873"},{"key":"2023101823050710000_2023.10.13.561998v2.5","doi-asserted-by":"publisher","DOI":"10.3390\/membranes13020250"},{"key":"2023101823050710000_2023.10.13.561998v2.6","doi-asserted-by":"publisher","DOI":"10.1038\/nature10545"},{"key":"2023101823050710000_2023.10.13.561998v2.7","doi-asserted-by":"publisher","DOI":"10.1038\/nrm.2017.16"},{"key":"2023101823050710000_2023.10.13.561998v2.8","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2023.104763"},{"key":"2023101823050710000_2023.10.13.561998v2.9","doi-asserted-by":"publisher","DOI":"10.1038\/s42003-022-03841-8"},{"key":"2023101823050710000_2023.10.13.561998v2.10","doi-asserted-by":"publisher","DOI":"10.1213\/ANE.0000000000004216"},{"key":"2023101823050710000_2023.10.13.561998v2.11","doi-asserted-by":"publisher","DOI":"10.1101\/2022.12.12.520161"},{"key":"2023101823050710000_2023.10.13.561998v2.12","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1407160111"},{"key":"2023101823050710000_2023.10.13.561998v2.13","doi-asserted-by":"crossref","unstructured":"Petersen, E.N. , Pavel, M.A. , Hansen, S.S. , Gudheti, M. , Wang, H. , Yuan, Z. , Murphy, K.R. , Ja, W.W. , Ferris, H.A. , Jorgensen, E.M. , et al.(2023). Mechanical activation of TWIK-related potassium channel by nanoscopic movement and rapid second messenger signaling. 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