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Using these methods, we have analyzed time-lapse fluorescence recordings of fluorescent keratin 13 in human vulva carcinoma-derived A431 cells. The fluorescent keratins integrated into the endogenous keratin cytoskeleton, and thereby served as reliable markers of keratin dynamics. We found that increased times after seeding correlated with down-regulation of inward-directed keratin filament movement. Bulk flow analyses further revealed that keratin filament polymerization in the cell periphery and keratin depolymerization in the more central cytoplasm were both reduced. Treating these cells and other human keratinocyte-derived cells with EGF reversed all these processes within a few minutes, coinciding with increased keratin phosphorylation. These results highlight the value of the newly developed tools for identifying modulators of keratin filament network dynamics and characterizing their mode of action, which, in turn, contributes to understanding the close link between keratin filament network plasticity and epithelial physiology.<\/jats:p>","DOI":"10.1073\/pnas.1306020110","type":"journal-article","created":{"date-parts":[[2013,6,11]],"date-time":"2013-06-11T21:14:26Z","timestamp":1370985266000},"page":"10664-10669","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":53,"title":["Measuring the regulation of keratin filament network dynamics"],"prefix":"10.1073","volume":"110","author":[{"given":"Marcin","family":"Moch","sequence":"first","affiliation":[{"name":"Institutes of aMolecular and Cellular Anatomy and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gerlind","family":"Herberich","sequence":"additional","affiliation":[{"name":"Imaging and Computer Vision, RWTH Aachen University, 52074 Aachen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Til","family":"Aach","sequence":"additional","affiliation":[{"name":"Imaging and Computer Vision, RWTH Aachen University, 52074 Aachen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rudolf E.","family":"Leube","sequence":"additional","affiliation":[{"name":"Institutes of aMolecular and Cellular Anatomy and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Reinhard","family":"Windoffer","sequence":"additional","affiliation":[{"name":"Institutes of aMolecular and Cellular Anatomy and"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2013,6,11]]},"reference":[{"key":"e_1_3_3_1_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ceb.2012.10.018"},{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.201008095"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.099655"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1172\/JCI37762"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.4161\/bioa.1.1.14815"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.068080"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.e03-09-0707"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.153.3.503"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.112.24.4521"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1002\/cm.1039"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1007\/s004410100455"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1523-1747.2003.12038.x"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.00096"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.105.3.753"},{"key":"e_1_3_3_15_2","first-page":"1359","article-title":"Fluorescence microscopic imaging and image analysis of the cytoskeleton","author":"Herberich G","year":"2010","unstructured":"G Herberich, et al., Fluorescence microscopic imaging and image analysis of the cytoskeleton. 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