{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T20:42:32Z","timestamp":1792183352015,"version":"4.4.0"},"reference-count":57,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2021,12,2]],"date-time":"2021-12-02T00:00:00Z","timestamp":1638403200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["390874280"],"award-info":[{"award-number":["390874280"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["INST 152\/772-1"],"award-info":[{"award-number":["INST 152\/772-1"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["152\/774-1"],"award-info":[{"award-number":["152\/774-1"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["152\/775-1"],"award-info":[{"award-number":["152\/775-1"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["152\/776-1"],"award-info":[{"award-number":["152\/776-1"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["152\/777-1 FUGG"],"award-info":[{"award-number":["152\/777-1 FUGG"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Leibniz ScienceCampus InterACt","award":["W6\/2018"],"award-info":[{"award-number":["W6\/2018"]}]},{"name":"Studienstiftung des Deutschen Volkes","award":["..."],"award-info":[{"award-number":["..."]}]},{"DOI":"10.13039\/100004410","name":"European Molecular Biology Organization","doi-asserted-by":"publisher","award":["ALTF356-2018"],"award-info":[{"award-number":["ALTF356-2018"]}],"id":[{"id":"10.13039\/100004410","id-type":"DOI","asserted-by":"publisher"}]},{"name":"HPI Strategic Incentive Programme","award":["..."],"award-info":[{"award-number":["..."]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The human adenovirus type 5 (HAdV5) infects epithelial cells of the upper and lower respiratory tract. The virus causes lysis of infected cells and thus enables spread of progeny virions to neighboring cells for the next round of infection. The mechanism of adenovirus virion egress across the nuclear barrier is not known. The human adenovirus death protein (ADP) facilitates the release of virions from infected cells and has been hypothesized to cause membrane damage. Here, we set out to answer whether ADP does indeed increase nuclear membrane damage. We analyzed the nuclear envelope morphology using a combination of fluorescence and state-of-the-art electron microscopy techniques, including serial block-face scanning electron microscopy and electron cryo-tomography of focused ion beam-milled cells. We report multiple destabilization phenotypes of the nuclear envelope in HAdV5 infection. These include reduction of lamin A\/C at the nuclear envelope, large-scale membrane invaginations, alterations in double membrane separation distance and small-scale membrane protrusions. Additionally, we measured increased nuclear membrane permeability and detected nuclear envelope lesions under cryoconditions. Unexpectedly, and in contrast to previous hypotheses, ADP did not have an effect on lamin A\/C reduction or nuclear permeability.<\/jats:p>","DOI":"10.3390\/ijms222313034","type":"journal-article","created":{"date-parts":[[2021,12,2]],"date-time":"2021-12-02T02:40:02Z","timestamp":1638412802000},"page":"13034","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Human Adenovirus Type 5 Infection Leads to Nuclear Envelope Destabilization and Membrane Permeability Independently of Adenovirus Death Protein"],"prefix":"10.3390","volume":"22","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0155-0317","authenticated-orcid":false,"given":"S\u00f8ren","family":"Pfitzner","sequence":"first","affiliation":[{"name":"Leibniz Institute for Experimental Virology (HPI), Martinistra\u00dfe 52, 20251 Hamburg, Germany"},{"name":"Centre for Structural Systems Biology, Notkestra\u00dfe 85, 22607 Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jens B.","family":"Bosse","sequence":"additional","affiliation":[{"name":"Leibniz Institute for Experimental Virology (HPI), Martinistra\u00dfe 52, 20251 Hamburg, Germany"},{"name":"Centre for Structural Systems Biology, Notkestra\u00dfe 85, 22607 Hamburg, Germany"},{"name":"Cluster of Excellence RESIST (EXC 2155), Hannover Medical School, Carl-Neuberg-Stra\u00dfe 1, 30625 Hannover, Germany"},{"name":"Hannover Medical School, Institute of Virology, Carl-Neuberg-Stra\u00dfe 1, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Helga","family":"Hofmann-Sieber","sequence":"additional","affiliation":[{"name":"Leibniz Institute for Experimental Virology (HPI), Martinistra\u00dfe 52, 20251 Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7691-0519","authenticated-orcid":false,"given":"Felix","family":"Flomm","sequence":"additional","affiliation":[{"name":"Leibniz Institute for Experimental Virology (HPI), Martinistra\u00dfe 52, 20251 Hamburg, Germany"},{"name":"Centre for Structural Systems Biology, Notkestra\u00dfe 85, 22607 Hamburg, Germany"},{"name":"Cluster of Excellence RESIST (EXC 2155), Hannover Medical School, Carl-Neuberg-Stra\u00dfe 1, 30625 Hannover, Germany"},{"name":"Hannover Medical School, Institute of Virology, Carl-Neuberg-Stra\u00dfe 1, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rudolph","family":"Reimer","sequence":"additional","affiliation":[{"name":"Leibniz Institute for Experimental Virology (HPI), Martinistra\u00dfe 52, 20251 Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Dobner","sequence":"additional","affiliation":[{"name":"Leibniz Institute for Experimental Virology (HPI), Martinistra\u00dfe 52, 20251 Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4788-2691","authenticated-orcid":false,"given":"Kay","family":"Gr\u00fcnewald","sequence":"additional","affiliation":[{"name":"Leibniz Institute for Experimental Virology (HPI), Martinistra\u00dfe 52, 20251 Hamburg, Germany"},{"name":"Centre for Structural Systems Biology, Notkestra\u00dfe 85, 22607 Hamburg, Germany"},{"name":"Universit\u00e4t Hamburg, Institute for Biochemistry and Molecular Biology, Martin-Luther-King-Platz 6, 20146 Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3040-8192","authenticated-orcid":false,"given":"Linda E.","family":"Franken","sequence":"additional","affiliation":[{"name":"Leibniz Institute for Experimental Virology (HPI), Martinistra\u00dfe 52, 20251 Hamburg, Germany"},{"name":"Centre for Structural Systems Biology, Notkestra\u00dfe 85, 22607 Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,12,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1006\/viro.1996.0295","article-title":"The E3-11.6-kDa adenovirus death protein (ADP) is required for efficient cell death: Characterization of cells infected with adp mutants","volume":"220","author":"Tollefson","year":"1996","journal-title":"Virology"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"12243","DOI":"10.1128\/JVI.78.22.12243-12251.2004","article-title":"Conditionally Replicating Adenoviruses Kill Tumor Cells via a Basic Apoptotic Machinery-Independent Mechanism That Resembles Necrosis-Like Programmed Cell Death","volume":"78","author":"Abbas","year":"2004","journal-title":"J. Virol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"3206","DOI":"10.1038\/s41419-017-0110-8","article-title":"RIPK3 promotes adenovirus type 5 activity","volume":"8","author":"Weigert","year":"2017","journal-title":"Cell Death Dis."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Raghava, S., Giorda, K.M., Romano, F.B., Heuck, A.P., and Hebert, D.N. (2011). The SV40 late protein VP4 is a viroporin that forms pores to disrupt membranes for viral release. PLoS Pathog., 7.","DOI":"10.1371\/journal.ppat.1002116"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"8285","DOI":"10.1128\/JVI.00741-11","article-title":"Nuclear Envelope Breakdown Can Substitute for Primary Envelopment-Mediated Nuclear Egress of Herpesviruses","volume":"85","author":"Klupp","year":"2011","journal-title":"J. Virol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2544","DOI":"10.1091\/mbc.e08-01-0009","article-title":"A Supraphysiological Nuclear Export Signal Is Required for Parvovirus Nuclear Export","volume":"219","author":"Engelsma","year":"2008","journal-title":"Mol. Biol. Cell"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"5697","DOI":"10.1128\/JVI.75.12.5697-5702.2001","article-title":"Herpes Simplex Virus Nucleocapsids Mature to Progeny Virions by an Envelopment \u2192 Deenvelopment \u2192 Reenvelopment Pathway","volume":"75","author":"Skepper","year":"2001","journal-title":"J. Virol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1692","DOI":"10.1016\/j.cell.2015.11.029","article-title":"Structural Basis of Vesicle Formation at the Inner Nuclear Membrane","volume":"163","author":"Hagen","year":"2015","journal-title":"Cell"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2296","DOI":"10.1128\/jvi.70.4.2296-2306.1996","article-title":"The adenovirus death protein (E3-11.6K) is required at very late stages of infection for efficient cell lysis and release of adenovirus from infected cells","volume":"70","author":"Tollefson","year":"1996","journal-title":"J. Virol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"743","DOI":"10.1016\/0042-6822(92)90250-S","article-title":"The E3-11. 6K Protein of adenovirus is an Asn-glycosylated integral membrane protein that localizes to the nuclear membrane","volume":"753","author":"Scaria","year":"1992","journal-title":"Virology"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1006\/viro.1998.9135","article-title":"Adenovirus death protein, a transmembrane protein encoded in the E3 region, is palmitoylated at the cytoplasmic tail","volume":"244","author":"Hausmann","year":"1998","journal-title":"Virology"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1006\/viro.2002.1772","article-title":"Overexpression of the ADP (E3-11.6K) protein increases cell lysis and spread of Adenovirus","volume":"305","author":"Doronin","year":"2003","journal-title":"Virology"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"240","DOI":"10.1016\/j.virol.2004.06.007","article-title":"Overexpression of adenovirus E3-11.6K protein induces cell killing by both caspase-dependent and caspase-independent mechanisms","volume":"326","author":"Zou","year":"2004","journal-title":"Virology"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1007\/BF01314784","article-title":"The kinetics of adenovirus infection and spread in cell cultures infected with low multiplicity","volume":"54","author":"Wigand","year":"1977","journal-title":"Arch. Virol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"10123","DOI":"10.1128\/JVI.01102-12","article-title":"Cell-Free Transmission of Human Adenovirus by Passive Mass Transfer in Cell Culture Simulated in a Computer Model","volume":"86","author":"Yakimovich","year":"2012","journal-title":"J. Virol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1861","DOI":"10.1002\/1873-3468.13848","article-title":"The Adenovirus Death Protein\u2014A small membrane protein controls cell lysis and disease","volume":"594","author":"Georgi","year":"2020","journal-title":"FEBS Lett."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"102543","DOI":"10.1016\/j.isci.2021.102543","article-title":"Microscopy deep learning predicts virus infections and reveals mechanics of lytic-infected cells","volume":"24","author":"Andriasyan","year":"2021","journal-title":"iScience"},{"key":"ref_18","first-page":"363","article-title":"Experimental adenovirus infection of the mouse adrenal gland. I. Light microscopic observation","volume":"75","author":"Hoenig","year":"1974","journal-title":"Am. J. Pathol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1016\/S0248-4900(98)80230-X","article-title":"Release of viruses and viral DNA from nucleus to cytoplasm of HeLa cells at late stages of productive adenovirus infection as revealed by electron microscope in situ hybridization","volume":"90","author":"Besse","year":"1998","journal-title":"Biol. Cell"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"e37","DOI":"10.1093\/nar\/gkt1339","article-title":"Improved seamless mutagenesis by recombineering using ccdB for counterselection","volume":"42","author":"Wang","year":"2014","journal-title":"Nucleic Acids Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1698","DOI":"10.1016\/j.celrep.2017.05.008","article-title":"An Engineered Virus Library as a Resource for the Spectrum-wide Exploration of Virus and Vector Diversity","volume":"19","author":"Zhang","year":"2017","journal-title":"Cell Rep."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"5087","DOI":"10.1073\/pnas.0710625105","article-title":"Mechanism and kinetics of pore formation in membranes by water-soluble amphipathic peptides","volume":"105","author":"Lee","year":"2008","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2510","DOI":"10.1074\/jbc.M112.428425","article-title":"The viroporin activity of the minor structural proteins VP2 and VP3 is required for SV40 propagation","volume":"288","author":"Giorda","year":"2013","journal-title":"J. Biol. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1038\/s41586-021-03819-2","article-title":"Highly accurate protein structure prediction with AlphaFold","volume":"596","author":"Jumper","year":"2021","journal-title":"Nature"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"7764","DOI":"10.1128\/JVI.77.14.7764-7778.2003","article-title":"Mutations within the ADP (E3-11.6K) protein alter processing and localization of ADP and the kinetics of cell lysis of adenovirus-infected cells","volume":"77","author":"Tollefson","year":"2003","journal-title":"J. Virol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"903","DOI":"10.1128\/JVI.01675-13","article-title":"Adenovirus death protein (ADP) is required for lytic infection of human lymphocytes","volume":"88","author":"Murali","year":"2014","journal-title":"J. Virol."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Pfitzner, S., Hofmann-Sieber, H., Bosse, J.B., Franken, L.E., Gr\u00fcnewald, K., and Dobner, T. (2020). Fluorescent protein tagging of adenoviral proteins pV and pIX reveals \u2018late virion accumulation compartment\u2019. PLoS Pathog., 16.","DOI":"10.1371\/journal.ppat.1008588"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1006\/jsbi.1996.0013","article-title":"Computer Visualization of Three-Dimensional Image Data Using IMOD","volume":"116","author":"Kremer","year":"1996","journal-title":"J. Struct. Biol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"e01002-20","DOI":"10.1128\/AAC.01002-20","article-title":"The FDA-Approved Drug Nelfinavir Inhibits Lytic Cell-Free but not Cell-Associated Nonlytic Transmission of Human Adenovirus","volume":"64","author":"Georgi","year":"2020","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Brandenburg, B., Lee, L.Y., Lakadamyali, M., Rust, M.J., Zhuang, X., and Hogle, J.M. (2007). Imaging poliovirus entry in live cells. PLoS Biol., 5.","DOI":"10.1371\/journal.pbio.0050183"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1053","DOI":"10.1038\/nature02836","article-title":"Structural rearrangements in the membrane penetration protein of a non-enveloped virus","volume":"26","author":"Dormitzer","year":"2004","journal-title":"Nature"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"9920","DOI":"10.1128\/JVI.76.19.9920-9933.2002","article-title":"Strategy for Nonenveloped Virus Entry: A Hydrophobic Conformer of the Reovirus Membrane Penetration Protein \u03bc1 Mediates Membrane Disruption","volume":"76","author":"Chandran","year":"2002","journal-title":"J. Virol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1992","DOI":"10.1128\/JVI.79.4.1992-2000.2005","article-title":"Adenovirus Protein VI Mediates Membrane Disruption following Capsid Disassembly","volume":"79","author":"Wiethoff","year":"2005","journal-title":"J. Virol."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Daniels, R., Sadowicz, D., and Hebert, D.N. (2007). A very late viral protein triggers the lytic release of SV40. PLoS Pathog., 3.","DOI":"10.1371\/journal.ppat.0030098"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Suzuki, T., Orba, Y., Okada, Y., Sunden, Y., Kimura, T., Tanaka, S., Nagashima, K., Hall, W.W., and Sawa, H. (2010). The human polyoma JC virus agnoprotein acts as a viroporin. PLoS Pathog., 6.","DOI":"10.1371\/journal.ppat.1000801"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Panou, M.M., Prescott, E.L., Hurdiss, D.L., Swinscoe, G., Hollinshead, M., Caller, L.G., Morgan, E.L., Carlisle, L., M\u00fcller, M., and Antoni, M. (2018). Agnoprotein is an essential egress factor during BK Polyomavirus infection. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19030902"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"6556","DOI":"10.1128\/JVI.00146-14","article-title":"Nuclear Magnetic Resonance Structure Revealed that the Human Polyomavirus JC Virus Agnoprotein Contains an \u03b1-Helix Encompassing the Leu\/Ile\/Phe-Rich Domain","volume":"88","author":"Coric","year":"2014","journal-title":"J. Virol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"210","DOI":"10.1016\/j.chom.2011.08.010","article-title":"Kinesin-1-mediated capsid disassembly and disruption of the nuclear pore complex promote virus infection","volume":"10","author":"Strunze","year":"2011","journal-title":"Cell Host Microbe"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"6009","DOI":"10.1128\/JVI.03531-14","article-title":"Influenza Virus-Induced Caspase-Dependent Enlargement of Nuclear Pores Promotes Nuclear Export of Viral Ribonucleoprotein Complexes","volume":"89","author":"Dzieciolowski","year":"2015","journal-title":"J. Virol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"370","DOI":"10.1172\/JCI200419670","article-title":"Lamin A\/C deficiency causes defective nuclear mechanics and mechanotransduction","volume":"113","author":"Lammerding","year":"2004","journal-title":"J. Clin. Investig."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1007\/s00395-010-0085-4","article-title":"Genetic and ultrastructural studies in dilated cardiomyopathy patients: A large deletion in the lamin A\/C gene is associated with cardiomyocyte nuclear envelope disruption","volume":"105","author":"Gupta","year":"2010","journal-title":"Basic Res. Cardiol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"10100","DOI":"10.1073\/pnas.1812622115","article-title":"Fibroblasts lacking nuclear lamins do not have nuclear blebs or protrusions but nevertheless have frequent nuclear membrane ruptures","volume":"115","author":"Chen","year":"2018","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"5686","DOI":"10.3390\/v7112900","article-title":"3D Analysis of HCMV Induced-Nuclear Membrane Structures by FIB\/SEM Tomography: Insight into an Unprecedented Membrane Morphology","volume":"7","author":"Villinger","year":"2015","journal-title":"Viruses"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1967","DOI":"10.1093\/emboj\/21.8.1967","article-title":"Caspase-6 gene disruption reveals a requirement for lamin A cleavage in apoptotic chromatin condensation","volume":"21","author":"Ruchaud","year":"2002","journal-title":"EMBO J."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"3507","DOI":"10.1128\/jvi.67.6.3507-3514.1993","article-title":"The adenovirus L3 23-kilodalton proteinase cleaves the amino-terminal head domain from cytokeratin 18 and disrupts the cytokeratin network of HeLa cells","volume":"67","author":"Chen","year":"1993","journal-title":"J. Virol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"6684","DOI":"10.1073\/pnas.0702158104","article-title":"Intranuclear targeting and nuclear export of the adenovirus E1B-55K protein are regulated by SUMO1 conjugation","volume":"104","author":"Kindsmuller","year":"2007","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"480","DOI":"10.1016\/0042-6822(83)90274-X","article-title":"Monoclonal Antibodies Which Recognize Native and Denatured Forms of the Adenovirus DNA-Binding Protein","volume":"128","author":"Reich","year":"1983","journal-title":"Virology"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1038\/nmeth.2019","article-title":"Fiji: An open-source platform for biological-image analysis","volume":"9","author":"Schindelin","year":"2012","journal-title":"Nat. Methods"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"424","DOI":"10.1083\/jcb.30.2.424","article-title":"A new staining method (OTO) for enhancing contrast of lipid-containing membranes and droplets in osmium tetroxide-fixed tissue with osmiophilic thiocarbohydrazide (TCH)","volume":"30","author":"Seligman","year":"1966","journal-title":"J. Cell Biol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1111\/j.1365-2818.1982.tb00362.x","article-title":"Resin development for electron microscopy and an analysis of embedding at low temperature","volume":"126","author":"Carlemalm","year":"1982","journal-title":"J. Microsc."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Flomm, F.J., Soh, T.K., Schneider, C., Britt, H.M., Thalassinos, K., Pfitzner, S., Reimer, R., Gr\u00fcnewald, K., and Bosse, J.B. (2021). Intermittent bulk release of human cytomegalovirus through multivesicular bodies. bioRxiv.","DOI":"10.1101\/2020.12.31.424954"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1111\/j.1365-2818.2007.01775.x","article-title":"A technique for improved focused ion beam milling of cryo-prepared life science specimens","volume":"226","author":"Hayles","year":"2007","journal-title":"J. Microsc."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"107389","DOI":"10.1016\/j.jsb.2019.09.006","article-title":"Mind the gap: Micro-expansion joints drastically decrease the bending of FIB-milled cryo-lamellae","volume":"208","author":"Wolff","year":"2019","journal-title":"J. Struct. Biol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1016\/j.jsb.2016.06.007","article-title":"Implementation of a cryo-electron tomography tilt-scheme optimized for high resolution subtomogram averaging","volume":"197","author":"Hagen","year":"2017","journal-title":"J. Struct. Biol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1016\/j.jsb.2005.07.007","article-title":"Automated electron microscope tomography using robust prediction of specimen movements","volume":"152","author":"Mastronarde","year":"2005","journal-title":"J. Struct. Biol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"519","DOI":"10.1016\/j.jsb.2012.09.006","article-title":"RELION: Implementation of a Bayesian approach to cryo-EM structure determination","volume":"180","author":"Scheres","year":"2012","journal-title":"J. Struct. Biol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1038\/nmeth.4193","article-title":"MotionCor2\u2014Anisotropic correction of beam-induced motion for improved cryo-electron microscopy","volume":"14","author":"Zheng","year":"2017","journal-title":"Nat. Methods"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/23\/13034\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:38:43Z","timestamp":1760168323000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/23\/13034"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,12,2]]},"references-count":57,"journal-issue":{"issue":"23","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["ijms222313034"],"URL":"https:\/\/doi.org\/10.3390\/ijms222313034","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,12,2]]}}}