{"corpus_id":9770597,"paper_sha":"537be709d4ac3cd999a5b49748cd07abd14f1bee","doi":"10.3892/ol.2015.2856","arxiv_id":null,"pmid":25663887,"pmcid":"4314999","mag_id":1979999713,"dblp_id":null,"acl_id":null,"title":"Immunofluorescence analysis of cytokeratin 8/18 staining is a sensitive assay for the detection of cell apoptosis","year":2015,"publication_date":"2015-01-07","venue":"Oncology Letters","journal":{"name":"Oncology Letters","pages":"1227 - 1230","volume":"9"},"journal_issn":null,"journal_title":null,"publication_types":["JournalArticle"],"pubmed_pub_types":["Journal Article"],"s2_fields_of_study":["Biology","Medicine"],"reference_count":24,"citation_count":11,"influential_citation_count":0,"is_open_access":true,"arxiv_categories":null,"arxiv_license":null,"arxiv_journal_ref":null,"mesh_headings":null,"chemicals":null,"comments_corrections":null,"source_flags":5,"s2_open_access_pdf_url":"https://www.spandidos-publications.com/10.3892/ol.2015.2856/download","s2_open_access_landing_url":"https://www.semanticscholar.org/paper/537be709d4ac3cd999a5b49748cd07abd14f1bee","s2_open_access_license":"CCBY","s2_open_access_status":"GOLD","pmc_open_access_pdf_url":null,"pmc_open_access_landing_url":null,"pmc_open_access_license":null,"pmc_open_access_status":null,"unpaywall_open_access_pdf_url":null,"unpaywall_open_access_landing_url":null,"unpaywall_open_access_license":null,"unpaywall_open_access_status":null,"abstract":"Apoptosis is one of the major types of programmed cell death. During this process, cells experience a series of morphological and biochemical changes. Flow cytometric analysis of Annexin V staining of cell surface phosphatidylserine, in combination with a DNA-staining dye to probe the permeability of the cell membrane, is an established method for detecting apoptosis. The present study aimed to show that the immunofluorescence analysis of cytokeratin (CK) 8/18 staining may provide a new and sensitive assay for the detection of apoptotic cells. Tumor cells were treated with 20 μM cisplatin to induce apoptosis. Following 12 and 24 h of cisplatin treatment, cells were collected and stained with 4′,6-diamidine-2′-phenylindole dihydrochloride (DAPI) and fluorescein-labeled anti-CK8/18 antibody. The apoptotic cells were subsequently examined by fluorescence microscopy. Annexin V-fluorescein isothiocyanate/propidium iodide staining followed by flow cytometric analysis confirmed that cisplatin was able to induce apoptosis in tumor cells. Immunofluorescence analysis demonstrated that apoptotic cells had a distinct CK8/18 staining pattern. In living cells, CK8/18 was uniformly distributed in the cytoplasm and cytosol; however in the apoptotic cells with a condensed and/or fragmented apoptotic nucleus (as identified by DAPI staining), fluorescein-labeled anti-CK8/18 antibody exhibited unusual punctate and/or bubbly staining in the cytosol. In the apoptotic cells that could not be identified by DAPI staining, fluorescein-labeled CK8/18 displayed polarized aggregated staining in the cytosol. These results indicate that fluorescein-conjugated CK8/18 may be a useful and sensitive indicator of cell apoptosis.","claims":[{"public_id":"cl_b3bfb51ab296de53286e5fd4d9c37f94","status":"active","text":"Apoptotic cells showed distinct cytokeratin 8/18 staining patterns, including punctate, bubbly, and polarized aggregated cytosolic staining, compared with the uniform cytoplasmic distribution seen in living cells.","confidence":0.95,"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/claims/cl_b3bfb51ab296de53286e5fd4d9c37f94"},{"public_id":"cl_ca7570d915c14cf9d0093c2fd12ef604","status":"active","text":"Cisplatin treatment induced apoptosis in tumor cells under the reported conditions.","confidence":0.93,"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous 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