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. 2020 Dec 17;10:22111. doi: 10.1038/s41598-020-79185-2

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© The Author(s) 2020

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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G6PD and H6PD Gene Expression after siRNA transfection. Representative images of western blot analysis of MCF-7 (a), MDA-MB-231 (b) and A549 (c) (run under the same experimental conditions for each cell line), reporting G6PD, H6PD and Actin expression. Relative densitometry analyses of G6PD and H6PD expression in MCF-7 (d,g), MDA-MB-231 (e,h) and A549 (f,i), under control condition (CTR, green), scramble siRNA (Scr, black), siRNA G6PD (red) and siRNA H6PD (blue). In all cell lines, scramble siRNA-transfected cells did not show significant variations in the levels of G6PD and H6PD with respect to control ones. G6PD siRNA delivery significantly reduced G6PD expression in MCF-7 (d), MDA-MB-231 (e) and A549 (f), while keeping unaltered H6PD expression. Similarly, H6PD siRNA transfection selectively reduced enzyme expression in MCF-7 (g), MDA-MB-231 (h) and A549 (i). Western blot analysis confirmed the efficiency and selectivity of gene silencing in all cancer cell models, documenting the absence of any interference between the trigger enzymes G6PD and H6PD. Data are expressed as % ± SD of respective control condition (n = 3). **p < 0.01, ***p < 0.001 vs corresponding scramble. The full-length gels are presented in Supplementary Fig. 1.