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. 2019 Jun 13;9:8609. doi: 10.1038/s41598-019-44950-5

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© The Author(s) 2019

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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SIAE expression is induced by stable tet-responsive RES256 cells. RES256 cells were cultured in the presence (+) and absence (−) of doxycycline (dox) for 48 hours and analysed by western blot. SIAE was expressed by two wild-type clones, SIAE 1 and SIAE 2, and by the catalytic mutant, S127A. Importantly only when the cells were cultured with doxycycline for 48 hours was SIAE expressed. Equal protein loading between samples from each clone are confirmed with beta actin. A further immuno-reactive band of approximately 32 kDa for SIAE was seen (Supplementary Fig. 3). The identity of this band is unknown. Less protein was loaded in the S127A samples due to protein concentration of the lysate. Image acquired using the Licor Odyssey Clx equipped with ImageStudio 5 software. Representative image of n = 2.