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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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Endogenous total ROS and O₂ ⁻ levels correspond to KIF1Bβ expression in neuroblastoma cells. (A) Immunoblot analysis of KIF1Bβ in neuroblastoma cell lines NB1, CHP212 and SK-N-SH. (B) Flow cytometric analysis of intracellular ROS with DCFDA (mean ± SD; n = 3; *P < 0.05). (C) Intracellular O₂ ⁻ measured with lucigenin-based chemiluminescence assay (mean ± SD; n = 3; ****P < 0.0001). (D) Immunoblot analysis of SK-N-SH cells stably transduced with lentivirus encoding shRNAs targeting KIF1Bβ (shKIF1Bβ #3 and #5) or control virus (shSCR) and selected with 1 µg/ml puromycin for 8 days. (E) Corresponding fold change in flow cytometric analysis of intracellular ROS and (F) Fold change in intracellular O₂ ⁻ determined using lucigenin-based chemiluminescence assay of SK-N-SH cells stably transduced with lentiviruses as indicated (mean ± SD; n = 3; *P < 0.05; ****P < 0.0001).