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. 2014 Sep 1;15(9):15456–15474. doi: 10.3390/ijms150915456

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© 2014 by the authors; licensee MDPI, Basel, Switzerland.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).

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Stretch-mediated activation of VEGF promoter activity. Dual luciferase reporter assays were used to screen VEGF promoter activation upon stretching. C-28/I2 chondrocytes were transfected with a VEGF-promoter specific reporter construct (pVEGF-KpnI), expressing inducible Firefly luciferase, and a constitutive active Renilla luciferase for normalization purposes. Cells were stretched (8%, 0.5 Hz, 12 h square waveforms (A) and the ratio of Firefly-to-Renilla signals is shown. The unstrained control (ctrl) is set to 100% (i.e., 1); HRE activity (B) was measured using a Hypoxia Responsive Element (HRE) reporter construct (pGL3-HRE). Unstrained controls (ctrl), stretched cells (strain). ** indicates p ≤ 0.01, n.s. = not significant; n = 5.