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We demonstrate that inhibiting cytoskeletal dynamics with pharmacological agents or by genetic manipulation prevents the degranulation of neutrophils and mitochondrial DNA release required for NET formation. Wiskott-Aldrich syndrome protein\u2013deficient neutrophils are unable to polymerize actin and exhibit a block in both degranulation and DNA release. Similarly, neutrophils with a genetic defect in NADPH oxidase fail to induce either actin and tubulin polymerization or NET formation on activation. Moreover, neutrophils deficient in glutaredoxin 1 (Grx1), an enzyme required for deglutathionylation of actin and tubulin, are unable to polymerize either cytoskeletal network and fail to degranulate or release DNA. Collectively, cytoskeletal dynamics are achieved as a balance between reactive oxygen species\u2013regulated effects on polymerization and glutathionylation on the one hand and the Grx1-mediated deglutathionylation that is required for NET formation on the other.<\/jats:p>","DOI":"10.1083\/jcb.201611168","type":"journal-article","created":{"date-parts":[[2017,11,17]],"date-time":"2017-11-17T14:15:14Z","timestamp":1510928114000},"page":"4073-4090","update-policy":"https:\/\/doi.org\/10.1083\/jcb.crossmarkpolicy","source":"Crossref","is-referenced-by-count":136,"title":["ROS and glutathionylation balance cytoskeletal dynamics in neutrophil extracellular trap formation"],"prefix":"10.1083","volume":"216","author":[{"given":"Darko","family":"Stojkov","sequence":"first","affiliation":[{"name":"Institute of Pharmacology, University of Bern, Bern, Switzerland 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2389-4997","authenticated-orcid":true,"given":"Poorya","family":"Amini","sequence":"additional","affiliation":[{"name":"Institute of Pharmacology, University of Bern, Bern, Switzerland 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kevin","family":"Oberson","sequence":"additional","affiliation":[{"name":"Institute of Pharmacology, University of Bern, Bern, Switzerland 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5992-8247","authenticated-orcid":true,"given":"Christiane","family":"Sokollik","sequence":"additional","affiliation":[{"name":"Unit of Pediatric Infectious Diseases, University Children\u2019s Hospital Bern, Bern, Switzerland 2"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrea","family":"Duppenthaler","sequence":"additional","affiliation":[{"name":"Unit of Pediatric Infectious Diseases, University Children\u2019s Hospital Bern, Bern, Switzerland 2"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9404-7736","authenticated-orcid":true,"given":"Hans-Uwe","family":"Simon","sequence":"additional","affiliation":[{"name":"Institute of Pharmacology, University of Bern, Bern, Switzerland 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9855-4305","authenticated-orcid":true,"given":"Shida","family":"Yousefi","sequence":"additional","affiliation":[{"name":"Institute of Pharmacology, University of Bern, Bern, Switzerland 1"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"291","published-online":{"date-parts":[[2017,11,17]]},"reference":[{"key":"2023072308581033100_bib1","doi-asserted-by":"publisher","first-page":"832","DOI":"10.1182\/blood-2003-07-2624","article-title":"Rac2 is critical for neutrophil primary granule exocytosis","volume":"104","author":"Abdel-Latif","year":"2004","journal-title":"Blood."},{"key":"2023072308581033100_bib2","doi-asserted-by":"publisher","first-page":"178","DOI":"10.1002\/eji.201545615","article-title":"NET formation can occur independently of RIPK3 and MLKL signaling","volume":"46","author":"Amini","year":"2016","journal-title":"Eur. 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