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. 2015 Jun 15;5:11413. doi: 10.1038/srep11413

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Copyright © 2015, Macmillan Publishers Limited

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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Total RNA was extracted from salivary glands at the indicated times following RRSV infection and subjected to quantitative real-time PCR analysis using a pair of primers to specifically amplify the RRSV major capsid protein gene, P8. The relative P8 transcript levels were calculated using N. lugens 18S rRNA as an internal control. A mixture of 100 salivary glands was used as one biological sample due to the very low quantity of salivary glands. Samples from each time point were tested in three biological replicates, and the mean value used to analyze the relative transcript levels.