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. 2014 Jun 27;9(6):e100669. doi: 10.1371/journal.pone.0100669

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© 2014 Khosravi et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Serum starved C3H10T1/2 were pre-treated with Wnt3a- or conditioned media for 16 hours and then treated with or without TNF-α (20 ng/ml) in the presence of dichlorobenzimidazole riboside (an inhibitor of mRNA transcription) for various intervals. Total RNA was isolated from cell lysates and subjected to real time PCR. Data were plotted as percent log remaining lysyl oxidase mRNA levels (normalized to 18 s rRNA mRNA levels) vs time. The TNF-α induced loss of lysyl oxidase mRNA stability was calculated from the relative slopes of the lines of best fit. Student's t-test statistical analyses were performed by comparing the slopes and intercepts of these two lines and showed a P value of 0.006 (GraphPad Prism 5). Data are means ± SD (n = 3 for each time point) and are from one of two representative experiments with the same outcome.