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. 2019 Dec 9;9:18652. doi: 10.1038/s41598-019-55197-5

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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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Knocking down SIK1 expression increases Na_V1.5-dependent cancer cell invasiveness. (A) Effect of silencing SIK1 (siSIK1) in MDA-MB-231 cells on the mRNA expression of SIK1, SCN5A and SNAI1 genes. Results were obtained from 4 independent RT-qPCR experiments, 24 h after siRNA transfection, and are expressed relatively to the results obtained when transfecting control irrelevant siRNA (siCTL). P values indicate significant differences (Mann-Whitney rank sum test) as compared to siCTL condition. (B) Effect of silencing SIK1 (siSIK1) on the invasiveness of MDA-MB-231 shCTL and shNa_V1.5 cells. Results are coming from 4 independent experiments and are expressed relatively to the effect of the siCTL condition in shCTL cells. P values indicate significant differences (Mann-Whitney rank sum test) as compared to siCTL condition in shCTL cells. NS stands for “no statistical difference” between siCTL and siSIK1 conditions in shNa_V1.5 cells. (C) Relative H⁺ efflux measurements induced by reintroduction of 130 mM NaCl in MDA-MB-231 shCTL cells transfected with either siCTL or siSIK1, after a NH₄Cl wash pulse-induced intracellular acidification in a Na⁺-free solution. Results are coming from 5 independent experiments and are expressed relatively to the efflux measured in the siCTL condition. P value indicates a significant difference (Mann-Whitney rank sum test) when compared to the siCTL condition. (D) Relative H+ efflux measurements induced by reintroduction of 130 mM NaCl in MDA-MB-231 shNa_V1.5 cells transfected with either siCTL or siSIK1, as in C. Results are coming from 5 independent experiments and are expressed relatively to the efflux measured in the siCTL condition. NS indicates “no statistical difference” when compared to the siCTL condition. (E) Effect of silencing SIK1 (siSIK1) in MCF-7 cells on the mRNA expression of SIK1, SCN5A and SNAI1 genes. Results were obtained from 7 independent RT-qPCR experiments, 24 h after siRNA transfection, and are expressed relatively to the results obtained when transfecting control irrelevant siRNA (siCTL). P values indicate significant differences (Mann-Whitney rank sum test) as compared to the siCTL condition. (F) Effect of silencing SIK1 (siSIK1) on the invasiveness of MCF-7 cells, in the presence of 30 µM tetrodotoxin (TTX) or not (CTL, vehicle). Results are coming from 7 independent experiments and are expressed relatively to the effect of the CTL condition in siCTL cells. P values indicate significant differences (Mann-Whitney rank sum test). NS stands for “no statistical difference” between CTL and TTX conditions in siCTL cells.