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. 2019 May 9;10:939. doi: 10.3389/fmicb.2019.00939

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Copyright © 2019 Wang, Wang, Zhang, Zeng, Zheng, Chen, Zhang and Wei.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Tubulin inhibitor suppressed RYSV infection in the CNS of viruliferous N. cincticeps. (A) The effect of the tubulin inhibitor colchicine on live N. cincticeps was determined. The CNSs from the two groups were dissected at 3, 6, and 8 days after the microinjection and immunolabeled with α-tubulin antibody (α-tubulin-F; green) and virus-rhodamine (V-R; red). Scale bars, 200 μm. (B) The relative transcript levels of the N and M genes in the different samples outlined in (A) were analyzed using RT-qPCR at 1, 3, 5, 7, 9, and 11 days post-microinjection. P-values were estimated using Tukey’s honest significant difference (HSD) test and data represent means ± SE from three biological repeats and statistical significance is indicated by asterisks, *, 0.01 < p < 0.01. (C) The accumulation of RYSV N and P proteins in the CNS of N. cincticeps were detected using an immunoblotting assay with N-specific and P-specific IgG antibodies at 8 days post-microinjection. Actin was used as a loading control and detected with β-actin antibodies.