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. 2010 Jun 24;38(20):7068–7078. doi: 10.1093/nar/gkq569

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© The Author(s) 2010. 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/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — IRES activity of mRev-erb α is dynamically regulated and phase dependent. (A) The IRES activity of mRev-erb α was differentially regulated and phase dependent, and was analyzed with mRNA transfection. The IRES activity of the truncated form was nearly constant. This result is representative of at least three independent experiments. The error bars represent the mean ± SEM of duplicate measurements. The labels on the x axis indicate mRNA transfection time-harvest time pairs of each mRNA reporter after dexamethasone synchronization. The significance of differences of IRES activities between different phases was determined by one-way analysis of variance (ANOVA). *P < 0.005 (B) Dynamics of the binding affinity of hnRNP Q to mRev-erb α 5′-UTR mRNA after synchronization was analyzed with the UV crosslinking assay. The loading amounts of cytosolic extracts were confirmed by western blotting of the 14-3-3 level in (C). The normalized band intensities are shown below the bands. The intensity at 12 h after synchronization was arbitrarily set as 100. (C) Dynamics of the levels of cytosolic PTB after synchronization as analyzed by western blotting. 14-3-3 protein was used as a loading control. The normalized band intensities are shown below the bands. The intensity at 12 h after synchronization was arbitrarily set as 100.