Fig. 5. Effect of calcitriol and paricalcitol on the expression of -catenin direct transcriptional target genes, cyclin D1, axin 2, and VCAN/versican, during HASMC. HASMCs are incubated for 24 h in a high (3.3 mmol/l) phosphate (HP) medium (calcification medium) alone or supplemented with either calcitriol 10 8 M (HP CTR) or paricalcitol 3·10 8 M (HP PC) alone or with the addition to culture medium (100 ng/ml) of commercially available DKK-1, a specific endogenous extracellular antagonist of the Wnt signaling. Cells incubated in normal phosphate (0.9 mmol/l) medium are used as controls. mRNA levels are analyzed by real-time RT-PCR technique. Expression of target gene mRNA is normalized to the expression of GAPDH. Bars are means SE (3 independent experiments; 6 repetitions in each experiment). *P 0.05 vs. control. $P 0.05 vs. HP. #P 0.05 vs. the same treatment without DKK-1.
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