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. 2015 Jul 21;39(12):1733–1741. doi: 10.1038/ijo.2015.121

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Copyright © 2015 Macmillan Publishers Limited

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/4.0/

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MEST depletion substitutes IBMX as inducer of human adipocyte differentiation. hMADS cells were transfected at confluence (d-2) with siRNAs targeting MEST (siMEST) or a control siRNA (siC). Adipocyte differentiation was induced 2 days later (d0). (a) Representative bright-field microscopy images (× 40 magnification) of cells at d3. (b–e) hMADS cells were stimulated to undergo adipocyte differentiation in suboptimal differentiation media lacking either dexamethasone (−Dex) or IBMX (-IBMX). (b) Triglyceride accumulation was visualised by Oil Red O staining at d9. (c) Quantification of intracellular triglycerides at d9 (n=4). (d and e) Quantitative real-time reverse transcription PCR (RT-qPCR) analysis of adipocyte marker gene expression (d9) for cells differentiated without Dex (d) or without IBMX (e). Gene expression was normalised to TBP and is presented relative to siC-transfected cells that were differentiated in complete differentiation medium (n=4). (c–e) *P<0.05, **P<0.01, ***P<0.001 versus siC-transfected cells exposed to the same differentiation cocktail.