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. 2017 Jun 8;7:3068. doi: 10.1038/s41598-017-03036-w

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Timecourse of IPNV infection of CHSE-214 cells. VP2/VP3 expression was determined by immunofluorescence after 4 (B), 6 (C), 8 (D) and 12 (E) h post infection (hpi) in CHSE-214 cells. Cells were propagated at 60–70% confluence in 24-well plates with round glass coverslips. IPNV was inoculated at a MOI of 1 for 1 h. After 4, 6, 8 and 12 h of incubation at 20 °C, cells were processed by indirect immunofluorescence (IFI) using mouse oligoclonal anti-VP2/VP3 IPNV antibody and Alexa 488-conjugated donkey anti-mouse antibody. Representative images, recorded with an Olympus Spinning Disk IX81 microscope, are shown (bar scale = 20 μm). At each incubation time, 250 cells were counted. Number of VP2/VP3-expressing cells is shown as a percentage, normalized to mock cells (F).