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. 2017 May 11;7:1749. doi: 10.1038/s41598-017-01866-2

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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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WISP1 silencing induces adipogenic differentiation. 3T3-F442A preadipocytes were transfected for 48 hours with siRNA control or siWISP1 (30 pmoles). Then, the influence of WISP1 knockdown on (a) endogenous WISP1 mRNA levels and (b) secreted WISP1 protein levels in the culture media was determined by real-time PCR and ELISA. (c) The effect of WISP1 knockdown on lipid accumulation was monitored by microscopic imaging after Oil Red O-staining. mRNA levels of PPARG, ADIPOQ, LPL, FABP4, CD36, DLK1 and CEBPD were measured by real-time PCR. The values indicate the changes for the indicated sample compared to the siRNA-control. (d) 3T3-F442A preadipocytes were incubated in MDI media or alternatively transfected with si-WISP1 (30 pmoles) in the presence or the absence of conditioned media from L cells expressing Wnt3a (Wnt3a-CM) for 96 hours. The differentiation process was monitored by microscopic imaging after Oil Red O-staining. Red-stained areas were quantified by ImageJ analysis. All values are averages of data from 3 independent experiments each performed in duplicate. Results are presented as means ± SEM; *p < 0.05; **p < 0.01; ***p < 0.005.