Figure 2

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.