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. 2019 Mar 21;11(3):285. doi: 10.3390/v11030285

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© 2019 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Knockdown of BSRF1 decreased progeny production. (A) Effect of BSRF1 knockdown on viral lytic protein expression in B95-8 cells. B95-8 cells were transfected by electroporation with two independent siRNAs (siBSRF1-1 and -2) or control siRNA and incubated. Cells were harvested after 3 days and subjected to immunoblotting with the indicated antibodies. (B,C) Infectivity of the progeny virus particles produced from B95-8. Six days after the transfection of siRNA as in (A), progeny virus produced into the media was inoculated with Akata(−) cells for 2 days, followed by immunofluorescence assay by using anti-LMP1 antibody (B). Percentage of green fluorescence-positive cells over total cells is shown in (C). Production of progeny in the culture media. B95-8 cells were transfected and cultured as in (B). (D) The medium was collected after centrifugation, treated with DNase, and subjected to DNA extraction and quantification by qPCR. The means ± SD of three independent biological replicates are shown. n.s, not significant.; *, P < 0.05; ***, P < 0.0001.