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High Content Image Based Analysis Identifies Cell Cycle Inhibitors as Regulators of Ebola Virus Infection
PerkinElmer, 940 Waltham, MA 02451, USA
United States Army Medical Research Institute of Infectious Diseases, 1425 Porter st, Fort Detrick, Frederick, MD 21702, USA
These authors contributed equally to this work.
* Author to whom correspondence should be addressed.
Received: 5 August 2012; in revised form: 9 September 2012 / Accepted: 11 September 2012 / Published: 25 September 2012
Abstract: 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.
Keywords: ebolavirus; cell cycle; high-content imaging; serum starvation; aphidicolin; nocodazole
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MDPI and ACS Style
Kota, K.P.; Benko, J.G.; Mudhasani, R.; Retterer, C.; Tran, J.P.; Bavari, S.; Panchal, R.G. High Content Image Based Analysis Identifies Cell Cycle Inhibitors as Regulators of Ebola Virus Infection. Viruses 2012, 4, 1865-1877.
Kota KP, Benko JG, Mudhasani R, Retterer C, Tran JP, Bavari S, Panchal RG. High Content Image Based Analysis Identifies Cell Cycle Inhibitors as Regulators of Ebola Virus Infection. Viruses. 2012; 4(10):1865-1877.
Kota, Krishna P.; Benko, Jacqueline G.; Mudhasani, Rajini; Retterer, Cary; Tran, Julie P.; Bavari, Sina; Panchal, Rekha G. 2012. "High Content Image Based Analysis Identifies Cell Cycle Inhibitors as Regulators of Ebola Virus Infection." Viruses 4, no. 10: 1865-1877.