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. 2018 Sep 28;8:14543. doi: 10.1038/s41598-018-32400-7

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© The Author(s) 2018

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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ZIKV infection of SH-SY5Y neuroblastoma cells following longer periods of retinoic acid-induced differentiation. (A) Infection protocol for undifferentiated cells (top) and cells after 2 to 8 days of differentiation with retinoic acid (bottom). Syringe icons denote the day of infection with ZIKV-UG. After one hour of incubation, virus was washed twice and collection of time 0 samples was performed. Infection was monitored after 48 hours by IFA, plaque assay and qRT-PCR. (B) Plot of viral titers as determined by plaque assay versus days of differentiation prior to ZIKV infection. (C) Plot of viral titers as determined by qRT-PCR at different days of differentiation. Gray bars represent control data from 0 hours post infection (hpi), and black bars represent data from 48 hpi. Titers and error bars are calculated from the average and standard deviation of three independent experiments. *p < 0.05 by analysis of variance (ANOVA); **p < 0.05 by one-tailed t-test; ***p = 0.07 by one-tailed t-test. Abbreviations: IFA, immunofluorescence assay; qRT-PCR, quantitative reverse transcription-polymerase chain reaction; hpi, hours post-infection; PFU, plaque-forming units.