PMR60 over-expression increases in vitro decay of PTH mRNA with an intact PTH mRNA 3'-UTR. IVDA using radiolabeled polyadenylated full-length rat PTH mRNA (top panel), GH mRNA (middle panel) or polyadenylated rat PTH mRNA with an internal deletion of the PTH mRNA 3'-UTR ARE (bottom panel) and extracts from HEK293 cells transfected with either PMR60, the catalytically inactive form, PMR600, or control pSG5 plasmid. A. Western blot analysis of protein extracts as above using anti Myc and β-actin antibodies, demonstrating equal amounts of Myc-PMR60 and Myc-PMR600 protein. B. RNA transcripts extracted at different time points, separated by agarose gels and visualized by autoradiography. C-E. Quantification of the IVDA results in A and in 2 repeat independent IVDA experiments, for PMR60 expressing or control extracts. Results are presented as mean ± S.E. of the amount of intact transcript at time 0 using a semi logarithmic scale. *, p < 0.05.
Nechama et al. BMC Cell Biology 2009 10:70 doi:10.1186/1471-2121-10-70