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. 2019 Nov 20;9:17152. doi: 10.1038/s41598-019-53676-3

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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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VIP induces the expression of glucose transporters GLUT1 and GLUT3 in human trophoblast-derived cells. (a,b) Cells were seeded until subconfluence and 50/100 nM VIP was added for 6 h in DMEM-F12 2% FBS and then cells were harvested. For mRNA analysis, qRT-PCR was performed and results were analysed employing 2^-ΔΔCT method normalized to the endogenous GAPDH gene control. For protein analysis, Western Blot was performed and GLUT1 expression was normalized to α-Tubulin or β-Actin. The image shows cropped lines corresponding to α-Tubulin, β-Actin and GLUT1. Blots were run under the same experimental conditions. Full-length gels are presented in Supplementary Fig. S1. Student’s t test or ANOVA with Dunnett’s multiple comparisons against basal condition was used to compare between conditions ((a) GLUT1 mRNA: n = 6 for Swan 71 and BeWo; GLUT1 protein: n = 3 for Swan 71 and BeWo; (b) GLUT3 mRNA: n = 5 for Swan 71 and n = 4 for BeWo). Results are expressed as Mean ± S.E.M. *p < 0.05, **p < 0.01.