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High Content Image Based Analysis Identifies Cell Cycle Inhibitors as Regulators of Ebola Virus Infection
Viruses modulate a number of host biological responses including the cell cycle to favor their replication. In this study, we developed a high-content imaging (HCI) assay to measure DNA content and identify different phases of the cell cycle. We then investigated the potential effects of cell cycle...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3497033/ https://www.ncbi.nlm.nih.gov/pubmed/23202445 http://dx.doi.org/10.3390/v4101865 |
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author | Kota, Krishna P. Benko, Jacqueline G. Mudhasani, Rajini Retterer, Cary Tran, Julie P. Bavari, Sina Panchal, Rekha G. |
author_facet | Kota, Krishna P. Benko, Jacqueline G. Mudhasani, Rajini Retterer, Cary Tran, Julie P. Bavari, Sina Panchal, Rekha G. |
author_sort | Kota, Krishna P. |
collection | PubMed |
description | Viruses modulate a number of host biological responses including the cell cycle to favor their replication. In this study, we developed a high-content imaging (HCI) assay to measure DNA content and identify different phases of the cell cycle. We then investigated the potential effects of cell cycle arrest on Ebola virus (EBOV) infection. Cells arrested in G1 phase by serum starvation or G1/S phase using aphidicolin or G2/M phase using nocodazole showed much reduced EBOV infection compared to the untreated control. Release of cells from serum starvation or aphidicolin block resulted in a time-dependent increase in the percentage of EBOV infected cells. The effect of EBOV infection on cell cycle progression was found to be cell-type dependent. Infection of asynchronous MCF-10A cells with EBOV resulted in a reduced number of cells in G2/M phase with concomitant increase of cells in G1 phase. However, these effects were not observed in HeLa or A549 cells. Together, our studies suggest that EBOV requires actively proliferating cells for efficient replication. Furthermore, multiplexing of HCI based assays to detect viral infection, cell cycle status and other phenotypic changes in a single cell population will provide useful information during screening campaigns using siRNA and small molecule therapeutics. |
format | Online Article Text |
id | pubmed-3497033 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-34970332012-11-29 High Content Image Based Analysis Identifies Cell Cycle Inhibitors as Regulators of Ebola Virus Infection Kota, Krishna P. Benko, Jacqueline G. Mudhasani, Rajini Retterer, Cary Tran, Julie P. Bavari, Sina Panchal, Rekha G. Viruses Article Viruses modulate a number of host biological responses including the cell cycle to favor their replication. In this study, we developed a high-content imaging (HCI) assay to measure DNA content and identify different phases of the cell cycle. We then investigated the potential effects of cell cycle arrest on Ebola virus (EBOV) infection. Cells arrested in G1 phase by serum starvation or G1/S phase using aphidicolin or G2/M phase using nocodazole showed much reduced EBOV infection compared to the untreated control. Release of cells from serum starvation or aphidicolin block resulted in a time-dependent increase in the percentage of EBOV infected cells. The effect of EBOV infection on cell cycle progression was found to be cell-type dependent. Infection of asynchronous MCF-10A cells with EBOV resulted in a reduced number of cells in G2/M phase with concomitant increase of cells in G1 phase. However, these effects were not observed in HeLa or A549 cells. Together, our studies suggest that EBOV requires actively proliferating cells for efficient replication. Furthermore, multiplexing of HCI based assays to detect viral infection, cell cycle status and other phenotypic changes in a single cell population will provide useful information during screening campaigns using siRNA and small molecule therapeutics. MDPI 2012-09-25 /pmc/articles/PMC3497033/ /pubmed/23202445 http://dx.doi.org/10.3390/v4101865 Text en © 2012 by the authors; licensee MDPI, Basel, Switzerland. http://creativecommons.org/licenses/by/3.0/ This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/). |
spellingShingle | Article Kota, Krishna P. Benko, Jacqueline G. Mudhasani, Rajini Retterer, Cary Tran, Julie P. Bavari, Sina Panchal, Rekha G. High Content Image Based Analysis Identifies Cell Cycle Inhibitors as Regulators of Ebola Virus Infection |
title | High Content Image Based Analysis Identifies Cell Cycle Inhibitors as Regulators of Ebola Virus Infection |
title_full | High Content Image Based Analysis Identifies Cell Cycle Inhibitors as Regulators of Ebola Virus Infection |
title_fullStr | High Content Image Based Analysis Identifies Cell Cycle Inhibitors as Regulators of Ebola Virus Infection |
title_full_unstemmed | High Content Image Based Analysis Identifies Cell Cycle Inhibitors as Regulators of Ebola Virus Infection |
title_short | High Content Image Based Analysis Identifies Cell Cycle Inhibitors as Regulators of Ebola Virus Infection |
title_sort | high content image based analysis identifies cell cycle inhibitors as regulators of ebola virus infection |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3497033/ https://www.ncbi.nlm.nih.gov/pubmed/23202445 http://dx.doi.org/10.3390/v4101865 |
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