Figure 2

The CaR decreases PTH mRNA levels and stability through the PTH mRNA 3'-UTR cis element. A. Effect of CaR over-expression on reporter GH mRNA containing the PTH mRNA ARE. qRT-PCR for GH and control HPRT mRNA levels in cells transfected with either control (CaR-) or CaR (CaR+) and expression plasmids for GH (left panel), GH containing the PTH mRNA 63 nt ARE (GH-PTH63) (middle panel) or GH containing a 40 nt truncated ARE (GH-tPTH40) (right panel). Results are fold changes compared with cells without the CaR expressed as mean ± SE of three experiments. *, P < 0.05. B. Effect of CaR on PTH mRNA decay. Representative IVDA of transcripts for the full-length hPTH mRNA and the PTH mRNA with an internal deletion of the 3'-UTR ARE, incubated with extracts from cells expressing either the CaR or empty vector. C. Quantification of the amount of intact transcripts remaining with time related to time 0 (mean ± SE, in three repeat experiments; *, P < 0.05). Blue square, full length PTH mRNA without CaR (CaR-) and red square, with CaR (CaR+); blue triangle, PTH w/o ARE without CaR (CaR-) and red triangle, with the CaR, (CaR+).