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. 2017 Mar 28;7:45375. doi: 10.1038/srep45375

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Copyright © 2017, The Author(s)

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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SK-N-SH cells were infected with JEV at an MOI of 1.0. The cells were collected at different time points. (A) Nedd4 mRNA levels were determined by RT-qPCR at different time points p.i. (B) The protein levels of Nedd4 and JEV NS3 were examined in mock- and JEV-infected SK-N-SH cells via Western blot using the indicated antibodies. GAPDH served as a control for equal sample loading. The level of Nedd4 was quantitated by densitometric analysis using ImageJ software and normalized to GAPDH. (C) Localization of Nedd4. SK-N-SH cells were fixed at 24 h p.i. and were subjected to immunofluorescence to detect JEV E (green) and Nedd4 (red). The nuclei were stained with DAPI (blue). Images were obtained using a confocal microscope (Scale bar = 10 μm). (D) The fluorescence intensity of Nedd4 was processed and quantified using ImageJ software. The data are shown as the mean ± SD of three independent experiments. *P < 0.05; **P < 0.01.