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. 2017 Dec 4;7:16867. doi: 10.1038/s41598-017-17192-6

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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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Apoptotic function of KIF1Bβ is necessary for ROS induction in neuroblastoma cells. (A) Immunoblot analysis of NB1 cells after 24 hours of transient transfection with 4 µg wild-type (FL) FLAG-KIF1Bβ or FLAG-KIF1Bβ mutants (E646V, P1217S, E1628K). Empty vector pcDNA3 (Empty) served as negative control. (B) Corresponding flow cytometric analysis of intracellular ROS in NB1 cells 24 hours post-transfection with FLAG-KIF1Bβ mutants as indicated. (C) Immunoblot analysis of NB1 cells after 24 hours of transient transfection with empty vector pcDNA3 (Empty) or FLAG-KIF1Bβ domain variants as indicated. (D) Corresponding fold change in flow cytometric analysis of intracellular ROS and (E) Fold change in intracellular O₂ ⁻ determined using lucigenin-based chemiluminescence assay 24 hours post-transfection in NB1 cells (mean ± SD; n = 3; *P < 0.05; **P < 0.01). (F) Immunoblot analysis of CHP212 cells after 24 hours of transient transfection with empty vector pcDNA3 (Empty) or FLAG-KIF1Bβ600-1400. (G) Corresponding fold change in flow cytometric analysis of intracellular ROS and (H) Fold change in intracellular O₂ ⁻ determined using lucigenin-based chemiluminescence assay 24 hours post-transfection in CHP212 cells (mean ± SD; n = 3; ****P < 0.0001).