The Expression of Clock and Clock-Controlled Genes Can Be Differentially Affected by HA-REV-ERBα Overexpression

<div><p>(A) TaqMan real-time RT-PCR of cDNA was performed with liver whole-cell RNA for the transcripts of <i>Dbp,</i> endogenous <i>Rev-erbα, mCry1, mCry2, mPer1,</i> and <i>mPer2</i> from untreated <i>LAP-tTA</i>/<i>TRE-Rev-erbα</i> mice (solid lines; −Dox) and Dox-treated <i>LAP-tTA</i>/<i>TRE-Rev-erbα</i> mice (dotted lines; +Dox).</p> <p>(B) Western blot analysis of liver nuclear extracts from <i>LAP-tTA</i>/<i>TRE-Rev-erbα</i> mice that were fed with normal chow (−Dox) or Dox-treated chow (+Dox). In accordance with the temporal mRNA profiles shown in (A) and (B), mCRY1 and mPER1 display reduced levels in untreated mice, whereas mPer2 and mCry2 accumulate to similar levels in nuclei of Dox-treated and untreated animals. The varying mPER1 migration is probably due to oscillating protein phosphorylation and dephosphorylation (see [<a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.0050034#pbio-0050034-b018" target="_blank">18</a>]).</p>