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. 2011 Dec 30;40(8):3419–3430. doi: 10.1093/nar/gkr1297

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© The Author(s) 2011. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 3. — Serum shock is sufficient to synchronize circadian gene expression in C2C12 myotubes. Stable clones of C2C12 cells (CE-4.7MyoD) were differentiated for 5 days prior to serum shock. RNA was isolated at the indicated time post-serum shock and used to determine the expression level of Bmal1 and MyoD by RT-PCR. The target transcript levels were normalized to Rpl26 transcript levels and then normalized to the highest point across all time points. The data for the graphs represent the mean values ± SEM for three independent clones for the reporter gene. The expression of endogenous Bmal1 (A), endogenous MyoD (B) all displayed a circadian oscillation as indicated by significant differences in peak and trough expression. For each respective gene, one-way ANOVA followed by Tukey post-hoc test identified significant difference (P < 0.05) between peak expression and other time points as indicated by an asterisk. For circadian analysis of gene expression the JTK_CYCLE program was used.