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. 2019 Jul 17;9:654. doi: 10.3389/fonc.2019.00654

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Copyright © 2019 Zheng, Wu, Zhang, Fu, Xu, Lao, Lin and Xu.

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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NBT causes G2/M arrest through up-regulation of GADD45α and the disruption of mitotic spindle formation. (A) Western blot analysis of mitosis-related proteins (cyclin B1 and GADD45α, GAPDH as loading control) after treatment of HeLa cells with 5 μM NBT. (B) Cells were treated with NBT (5 μM) for 8 h. Relative GADD45α mRNA levels (normalized to 18S mRNA) were analyzed by quantitative RT-PCR. The data shown are the means ± SD from 3 independent experiments and were analyzed using Student's t-test (***p < 0.001). (C) HeLa cells pre-treated with nocodazole (100 ng/ml) for 18 h were released into fresh medium or into medium containing NBT at the indicated times. Samples were analyzed by western blotting. (D) HeLa cells transfected with YFP-tubulin (green) were treated with nocodazole as in (C), Cells were stained with DAPI (blue) and analyzed by confocal microscopy (Leica TCS SP8, 100x oil). Scale bars: 25 μm. Noc. Rel. (nocodazole release).